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A process for the resolution of a racemic mixture of nucleoside enantiomers that includes the step of exposing the racemic mixture to an enzyme that preferentially catalyzes a reaction in one of the enantiomers. The nucleoside enantiomer (-)-2-hydroxymethyl-5-(5-fluorocytosin-1-yl)-1,3-oxathiolane is an effective antiviral agent against HIV, HBV, and other viruses replicating in a similar manner.
1. A method for the resolution of a racemic mixture of β-2-hydroxymethal-5-(cytosin-1-yl)-1,3-oxathiolane, comprising the step of selectively hydrolzing a 5'-O-acyl derivative of the racemic mixture of the nucleoside with an enzyme selected from the group consisting of pig liver esterase, porcine pancreatic lipase, and subtilisin.
2. The method of claim 1, wherein the enzyme is pig liver esterase.
3. The method of claim 1, wherein the enzyme is porcine pancreatic lipase.
4. The method of claim 1, wherein the enzyme is subtilisin.
5. The method of claim 1, wherein the 5'-O-acyl group is the residue of an alkyl carboxylic acid selected from the group consisting of acetic acid, propionic acid, butyric acid, and pentanoic acid.
6. A method for the resolution of a racemic mixture of β-2',3'-dideoxy-3'-thia-5fluorocytidine comprising the step of selectively hydrolyzing a 5'-O-acyl derivative of the racemic mixture of the nucleoside with an enzyme selected from the group consisting of pig liver esterase, porcine pancreatic lipase, and subtilisin.
7. The method of claim 6, wherein the enzyme is pig liver esterase.
8. The method of claim 6, wherein the enzyme is porcine pancreatic lipase.
9. The method of claim 6, wherein the enzyme is subtilisin.
10. The method of claim 1, wherein the 5'-O-acyl group is the residue of an alkyl carboxylic acid selected from the group consisting of acetic acid, propionic acid, butyric acid, and pentanoic acid.
The U.S. Government has rights in this invention arising out of the partial funding of work leading to this invention through the National Institutes of Health Grant Nos. NIH 5-21935 and NIH AI-26055, as well as a Veteran's Administration Merit Review Award.
This application is a continuation of U.S. Ser. No. 08/402,730 filed on Mar. 13, 1995, which is a continuation of U.S. Ser. No. 08/092,248 filed on Jul. 15, 1993 (now abandoned), which is a continuation of U.S. Ser. No. 07/736,089 filed on Jul. 26, 1991 (now abandoned), which is a continuation-in-part of U.S. Ser. No. 07/659,760 filed on Feb. 22, 1991 (now U.S. Pat. No. 5,210,085) which is a continuation-in-part of U.S. Ser. No. 07/473,318 filed on Feb. 1, 1990 (now U.S. Pat. No. 5,204,466), the contents of which are incorporated herein by reference.
This invention is in the area of biologically active nucleosides, and specifically includes a method for the resolution of nucleoside enantiomers, including 1,3-oxathiolane nucleosides, and antiviral compositions that include the enantiomerically enriched 1,3-oxathiolane nucleosides, (-) and (+)-2-hydroxymethyl-5-(5-fluorocytosin-l-yl)-1,3-oxathiolane ("FTC").
In 1981, acquired immune deficiency syndrome (AIDS) was identified as a disease that severely compromises the human immune system, and that almost without exception leads to death. In 1983, the etiological cause of AIDS was determined to be the human immunodeficiency virus (HIV). In December 1990, the World Health Organization estimated that between 8 and 10 million people worldwide were infected with HIV, and of that number, between 1,000,000 and 1,400,000 were in the U.S.
In 1985, it was reported that the synthetic nucleoside 3'-azido-3'-deoxythymidine (AZT) inhibits the replication of human immunodeficiency virus type 1. Since then, a number of other synthetic nucleosides, including 2',3'-dideoxyinosine (DDI), 2',3'-dideoxycytidine (DDC), 3'-fluoro-'3-deoxythymidine (FLT), 2',3'-dideoxy-2',3'-didehydrothymidine (D4T), and 3'-azido-2',3'-dideoxyuridine (AZDU), have been proven to be effective against HIV. A number of other 2',3'-dideoxynucleosides have been demonstrated to inhibit the growth of a variety of other viruses in vitro. It appears that, after cellular phosphorylation to the 5'-triphosphate by cellular kinases, these synthetic nucleosides are incorporated into a growing strand of viral DNA, causing chain termination due to the absence of the 3'-hydroxyl group.
In its triphosphate form, 3'-azido-3'-deoxythymidine is a potent inhibitor of HIV reverse transcriptase and has been approved by the FDA for the treatment of AIDS. However, the benefits of AZT must be weighed against the severe adverse reactions of bone marrow suppression, nausea, myalgia, insomnia, severe headaches, anemia, peripheral neuropathy, and seizures. These adverse side effects often occur immediately after treatment begins, whereas a minimum of six weeks of therapy is necessary to realize AZT's benefits. DDI, which has recently been approved by an FDA Committee for the treatment of AIDS, is also associated with a number of side effects, including sporadic pancreatis and peripheral neuropathy.
Both DDC and D4T are potent inhibitors of HIV replication with activities comparable (D4T) or superior (DDC) to AZT. However, both DDC and D4T are not efficiently converted to the corresponding 5'-triphosphates in vivo. Both compounds are also toxic and can cause peripheral neuropathies in humans.
The success of various 2',3'-dideoxynucleosides in inhibiting the replication of HIV in vivo or in vitro has led a number of researchers to design and test nucleosides that substitute a heteroatom for the carbon atom at the 3'-position of the nucleoside. Norbeck, et al., disclose that (▒)-1- (2β4β)-2-(hydroxymethyl)-4-dioxolanyl!thymine (referred to below as (▒)-dioxolane-T) exhibits a modest activity against HIV (EC50 of 20 μin ATH8 cells), and is not toxic to uninfected control cells at a concentration of 200μ. Tetrahedron Letters 30 (46), 6246, (1989).
European Patent Application Publication No. 0 382 526 filed by IAF Biochem International, Inc. discloses a number of substituted 1,3-oxathiolanes with antiviral activity, and specifically reports that the racemic mixture (about the C4'-position) of the C1'-B isomer of 2-hydroxymethyl-5-(cytosin-1-yl)-1,3-oxathiolane (referred to below as (▒)-BCH-189) has approximately the same activity against HIV as AZT, and no cellular toxicity at therapeutic levels. (▒)-BCH-189 has also been found to inhibit the replication of AZT-resistant, HIV isolates from patients who have been treated with AZT for longer than 36 weeks.
To market a nucleoside for pharmaceutical purposes, it must not only be efficacious with low toxicity, it must also be cost effective to manufacture. An extensive amount of research and development has been directed toward new, low cost processes for large scale nucleoside production. 2',3'-Dideoxynucleosides are currently prepared by either of two routes: derivatization of an intact nucleoside or condensation of a derivatized sugar moiety with a heterocyclic base. Although there are numerous disadvantages associated with obtaining new nucleoside analogues by modifying intact nucleosides, a major advantage of this approach is that the appropriate absolute stereochemistry has already been set by nature. However, this approach cannot be used in the production of nucleosides that contain either nonnaturally occurring bases or nonnaturally occurring carbohydrate moieties (and which therefore are not prepared from intact nucleosides), such as 1,3-oxathiolane nucleosides and 1,3-dioxolane nucleosides.
When condensing a carbohydrate or carbohydrate-like moiety with a heterocyclic base to form a synthetic nucleoside, a nucleoside is produced that has two chiral centers (at the C1' and C4'-positions), and thus exists as a diasteromeric pair. Each diastereomer exists as a set of enantiomers. Therefore, the product is a mixture of four enantiomers.
It is often found that nucleosides with nonnaturally- occurring stereochemistry in either the C1' or the C4'-positions are less active than the same nucleoside with the stereochemistry as set by nature. For example, Carter, et al., have reported that the concentration of the (-)-enantiomer of carbovir (2',3'-didehydro-2',3'-dideoxyguanosine) required to reduce the reverse transcriptase activity by 50% (EC50) is 0.8 μM, whereas the EC50) for the (+)-enantiomer of carbovir is greater than 60 μM. Antimicrobial Agents and Chemotherapy, 34:6, 1297-1300 (June 1990).
U.S. Ser No. 07/659,760 discloses that 1,3-oxathiolane and 1,3-dioxolane nucleosides can be prepared with high diastereoselectivity (high percentage of nucleoside with a B configuration of the bond from the C1'-carbon to the heterocyclic base) by careful selection of the Lewis acid used in the condensation process. It was discovered that condensation of a 1,3-oxathiolane nucleoside with a base occurs with almost complete β-stereospecificity when stannic chloride is used as the condensation catalyst, and condensation of 1,3-dioxolane with a base occurs with almost complete β-stereospecificity when various chlorotitanium catalysts are employed. Other Lewis acids provide low (or no) C1'-β selectivity or simply fail to catalyze the reactions.
There remains a strong need to provide a cost effective, commercially viable method to obtain β-stereospecificity of synthetic nucleosides prepared by condensing a carbohydrate-like moiety with a base. This is important because it is likely that many synthetic nucleoside inhibitors of viral replication now emerging from academic and commercial laboratories will require resolution. An economical and facile method for resolving racemic mixtures of nucleosides would greatly facilitate antiviral research and ultimately, commercial manufacture. Further, resolution of racemic mixtures of nucleosides may provide a route to increase the activity of synthetic nucleosides by eliminating or minimizing the undesired enantiomer.
Therefore, it is an object of the present invention to provide a method for the resolution of racemic mixtures of nucleosides.
It is another object of the present invention to provide enantiomerically enriched 1,3-oxathiolane nucleosides.
It is still another object of the present invention to provide enantiomerically enriched 1,3-oxathiolane nucleosides with significant antiviral activity and low toxicity.
A process for the resolution of a racemic mixture of nucleoside enantiomers or their derivatives is disclosed that includes the step of exposing the racemic mixture to an enzyme that preferentially catalyzes a reaction in one of the enantiomers. The process can be used to resolve a wide variety of nucleosides, including pyrimidine and purine nucleosides that are optionally substituted in the carbohydrate moiety or base moiety. The process can also be used to resolve nucleoside derivatives that contain additional heteroatoms in the carbohydrate moiety, for example, FTC and BCH-189. The resolution of nucleosides can be performed on large scale at moderate cost.
It has been discovered that the nucleoside enantiomer (-)-2-hydroxymethyl-5-(5-fluorocytosin-l-yl)-l,3-oxathiolane ("FTC") exhibits significant activity against HIV (EC50 of 0.0077 to 0.02 μM), HBV (hepatitis B virus), and other viruses replicating in a similar manner. The (+)-enantiomer of FTC is also active against HIV (EC50 of 0.28-0.84 μM).
FIG. 1 is an illustration of the chemical structure of 2-hydroxymethyl-5-(5-fluorocytosin-l-yl)-1,3-oxathiolane ("FTC").
FIG. 2 is an illustration of a method for the preparation of enantiomerically enriched cytidine and uridine 1,3-oxathiolane nucleosides.
FIG. 3 is a graph indicating the progress of lipase-catalyzed hydrolysis of the 5'-butyryl ester of FTC over time using the enzymes PS800 (-open square-) and PLE (-open circle with dot-).
FIG. 4 is a graph of the effect of concentration (μM) of racemic and enantiomerically enriched FTC (prepared by the method of Example 3) versus the percent inhibition of human PBM cells infected with HIV-1. ((-darkened circle-, (▒)-FTC), (-circle- (-)-FTC), (-darkened square-,(+)-FTC).
FIG. 5 is a graph of the effect of concentration (μM) of racemic and enantiomerically enriched FTC (prepared by method of Example 2) on the percent inhibition of human PBM cells infected with HIV-1. ((-darkened circle-, (▒)-FTC), (-circle -,(--)-FTC), (-darkened square-,(+)-FTC).
FIG. 6 is a graph of the effect of concentration (μM) of racemic and enantiomerically enriched BCH-189 (prepared by the method of Example 3) on the percent inhibition of human PBM cells infected with HIV-1. ((-darkened circle-, (▒)-BCH-189), (-circle-,(-)-BCH-189), (-darkened square-,(+)-BCH-189).
FIG. 7 is a graph of the uptake of tritiated (▒)-FTC in human PBM cells (average of two determinations) in time (hours) versus pmol/106 cells.
FIG. 8 is a graph of the egress of radiolabeled (▒)-FTC from human PBM cells, measured in hours versus pmol/106 cells ((-darkened square-), (▒)-FTC; (-///-), (▒)-FTC monophosphate; (-//-), (▒)-FTC diphosphate; and (-//////-), (▒)-FTC triphosphate).
As used herein, the term "enantiomerically enriched nucleoside" refers to a nucleoside composition that includes at least 95% of a single enantiomer of that nucleoside.
As used herein, the term BCH-189 refers to 2-hydroxymethyl-5-(cytosin-1-yl)-1,3-oxathiolane.
As used herein, the term FTC refers to 2-hydroxymethyl-5-(5-fluorocytosin-1-yl)-1,3-oxathiolane.
As used herein, the term "preferential enzyme catalysis" refers to catalysis by an enzyme that favors one substrate over another.
I. Resolution of Racemic Mixtures of Nucleosides During the Hydrolysis
A method is provided for the resolution of racemic mixtures of nucleoside enantiomers. The method involves the use of an enzyme that preferentially catalyzes a reaction of one enantiomer in a racemic mixture. The reacted enantiomer is separated from the unreacted enantiomer on the basis of the new difference in physical structure. Given the disclosure herein, one of skill in the art will be able to choose an enzyme that is selective for the nucleoside enantiomer of choice (or selective for the undesired enantiomer, as a method of eliminating it), by selecting of one of the enzymes discussed below or by systematic evaluation of other known enzymes. Given this disclosure, one of skill in the art will also know how to modify the substrate as necessary to attain the desired resolution. Through the use of either chiral NMR shift reagents, polarimetry, or chiral HPLC, the optical enrichment of the recovered ester can be determined.
The following examples further illustrate the use of enzymes to resolve racemic mixtures of enantiomers. Other known methods of resolution of racemic mixtures can be used in combination with the method of resolution disclosed herein. All of these modifications are considered within the scope of the invention.
Resolution Based on Hydrolysis of C5'-Nucleoside Esters
In one embodiment, the method includes reacting the C5'-hydroxyl group of a mixture of nucleoside racemates with an acyl compound to form C5'-esters in which the nucleoside is in the "carbinol" end of the ester. The racemic mixture of nucleoside C5'-esters is then treated with an enzyme that preferentially cleaves, or hydrolyses, one of the enantiomers and not the other.
An advantage of this method is that it can be used to resolve a wide variety of nucleosides, including pyrimidine and purine nucleosides that are optionally substituted in the carbohydrate moiety or base moiety. The method can also be used to resolve nucleoside derivatives that contain additional heteroatoms in the carbohydrate moiety, for example, FTC and BCH-189. The broad applicability of this method resides in part on the fact that although the carbinol portion of the ester plays a role in the ability of an enzyme to differentiate enantiomers, the major recognition site for these enzymes is in the carboxylic acid portion of the ester. Further, one may be able to successfully extrapolate the results of one enzyme/substrate study to another, seemingly-different system, provided that the carboxylic acid portions of the two substrates are the same or substantially similar.
Another advantage of this method is that it is regioselective. Enzymes that hydrolyse esters typically do not catalyze extraneous reactions in other portions of the molecule. For example, the enzyme lipase catalyses the hydrolysis of the ester of 2-hydroxymethyl-5-oxo-1,3-oxathiolane without hydrolysing the internal lactone. This contrasts markedly with "chemical" approaches to ester hydrolysis.
Still another advantage of this method is that the separation of the unhydrolysed enantiomer and the hydrolysed enantiomer from the reaction mixture is quite simple. The unhydrolysed enantiomer is more lipophilic than the hydrolysed enantiomer and can be efficiently recovered by simple extraction with one of a wide variety of nonpolar organic solvents or solvent mixtures, including hexane and hexane/ether. The less lipophilic, more polar hydrolysed enantiomer can then be obtained by extraction with a more polar organic solvent, for example, ethyl acetate, or by lyophilization, followed by extraction with ethanol or methanol. Alcohol should be avoided during the hydrolysis because it can denature enzymes under certain conditions.
Enzymes and Substrates
With the proper matching of enzyme and substrate, conditions can be established for the isolation of either nucleoside enantiomer. The desired enantiomer can be isolated by treatment of the racemic mixture with an enzyme that hydrolyses the desired enantiomer (followed by extraction of the polar hydrolysate with a polar solvent) or by treatment with an enzyme that hydrolyses the undesired enantiomer (followed by removal of the undesired enantiomer with a nonpolar solvent).
Enzymes that catalyze the hydrolysis of esters include esterases, for example pig liver esterase, lipases, including porcine pancreatic lipase and Amano PS-800 lipase, substillisin, and α-chymotrypsin.
The most effective acyl group to be used to esterify the C5'-position of the nucleoside can be determined without undue experimentation by evaluation of a number of homologs using the selected enzyme system. For example, when 1,3-oxathiolane nucleosides are esterified with butyric acid, resolutions with both pig liver esterase and Amano PS-800 proceed with high enantioselectivity (94-100 enantiomeric excess) and opposite selectivity. Non-limiting examples of other acyl groups that can be evaluated for use with a particular nucleoside enantiomeric mixture and particular enzyme include alkyl carboxylic acids and. substituted alkyl carboxylic acids, including acetic acid, propionic acid, butyric acid, and pentanoic acid. With certain enzymes, it may be preferred to use an acyl compound that is significantly electron-withdrawing to facilitate hydrolysis by weakening the ester bond. Examples of electron-withdrawing acyl groups include α-haloesters such as 2-chloropropionic acid, 2-chlorobutyric acid, and 2-chloropentanoic acid. α-Haloesters are excellent substrates for lipases.
Resolution Conditions
The enzymatic hydrolyses are typically carried out with a catalytic amount of the enzyme in an aqueous buffer that has a pH that is close to the optimum pH for the enzyme in question. As the reaction proceeds, the pH drops as a result of liberated carboxylic acid. Aqueous base should be added to maintain the pH near the optimum value for the enzyme. The progress of the reaction can be easily determined by monitoring the change in pH and the amount of base needed to maintain pH. The hydrophobic ester (the unhydrolysed enantiomer) and the more polar alcohol (the hydrolysed enantiomer) can be sequentially and selectively extracted from the solution by the judicious choice of organic solvents. Alternatively, the material to be resolved can be passed through a column that contains the enzyme immobilized on a solid support.
Enzymatic hydrolyses performed under heterogeneous conditions can suffer from poor reproducibility. Therefore, it is preferred that the hydrolysis be performed under homogeneous conditions. Alcohol solvents are not preferred because they can denature the enzymes. Homogeneity can be achieved through the use of non-ionic surfactants, such as Triton X-100. However, addition of these surfactants not only assists in dissolving the starting material, they also enhances the aqueous solubility of the product. Therefore, although the enzymatic reaction can proceed more effectively with the addition of a non-ionic surfactant than under heterogeneous conditions, the isolation of both the recovered starting material and the product can be made more difficult. The product can be isolated with appropriate chromatographic and chemical (e.g., selective salt formation) techniques. Diacylated nucleosides can be used but are often quite lipophilic and hard to dissolve in the medium used.
A number of 5'-O-acyl derivatives of FTC were prepared by selective O-acylation of the N-hydrochloride salt (see Table 1) of (▒)-FTC. The efficiency of the hydrolysis of the derivatives by lipases was investigated. As shown in Table 1, pig liver esterase (PLE) exhibits a high level of selectivity for the hydrolysis of the ester of the (+)-enantiomer of FTC. In contrast, PS-800 hydrolyses the ester of the (-)-enantiomer of FTC preferentially. The rate of the hydrolysis was also found to be dependent on the nature of the acyl group; the acetyl derivative was significantly slower than the butyryl derivative. It has now been discovered that the rate of the hydrolysis of the propionic acid ester of FTC is even faster than that observed for the butyrate derivative. % recovery and % ee were both determined using HPLC. Although the enantioselectivity is excellent when employing PLE (typically 97% e.e. or higher), additional enrichment can be accomplished by sequential enzymatic hydrolysis reactions in which the enantiomerically-enriched butyrate from a PLE-catalyzed hydrolysis is subjected to enzymatic hydrolysis by PS-800. Table 1: Comparison of Effect of Ester on Enzyme Hydrolysis .
TABLE 1______________________________________Comparison of Effect of Ester on Enzyme Hydrolysis.substrate % recovery % e.e. (s.m.)______________________________________FTC Esters with PLE:acetate 32.68 N.D.propionate 39.87 N.D.butyrate 48.00 98butyrate 45.71 98.6FTC Esters with PS800:acetate 73.17 N.D.propionate 52.67 N.D.butyrate 58.34 N.D.valerate 41.50 94______________________________________
The 5'-0-butyrate of (▒)-FTC (1) (0.47 mmol, 149 mg) was dissolved in 16 mL of a solution of 4:1 pH 8 buffer:CH3 CN. The clear solution was stirred and treated with 26 mg of pig liver esterase (PLE-A). The progress of the reaction was monitored by HPLC (FIG. 3). After 20 hours (52% conversion), the reaction mixture was extracted with 2ū80 mL of CHCl3 and 80 mL of ethyl acetate. The organic layer extracts were combined, dried over anhydrous MgSO4, filtered, and concentrated by rotary evaporation. The resulting residue was eluted on 2ū1000m pTLC plates using ethyl acetate as eluant (double elution) to give, after isolation, 53 mg (36% based on starting material) of FTC butyrate which was determined to have 98% enantiomeric excess (e.e.) by HPLC analysis. The enantiomerically-enriched butyrate was then treated with 1.6 mL of methanol followed by 0.38 mmol (20 mg) of sodium methoxide. The resulting mixture was stirred at room temperature, and the progress of the reaction was monitored by HPLC. The reaction was completed within 30 minutes. The solvent was removed by rotary evaporation to give a crude white solid (76 mg) that was eluted on a 1000 m pTLC using 5:1 ethyl acetate:ethanol. (-)-FTC was isolated as a white solid (33 mg; 82% yield). HPLC analysis of the FTC as its 5'-O-acetate derivative showed 97% e.e.; α!(20,D -120.5░(c=0.88; abs. ethanol).
Similarly, 1.2 mmol (375 mg) of the 5'-O-butyrate of (▒)-FTC was dissolved in 40 mL of 4:1 pH 8 buffer-CH3 CN. The clear solution was stirred and treated with 58 mg of pig liver esterase (PLE-A). The progress of the reaction was monitored by HPLC. After 90 minutes (38% conversion), the reaction mixture was added to 150 mL of CHCl3. The layers were separated and the aqueous layer lyophilized to remove solvent. The white residue from the lyophilization was extracted with 3ū10 mL of absolute ethanol. The extracts were filtered, combined, and concentrated in vacuo to yield 179 mg of crude oil. The crude material was eluted on a 45ū30 mm column of silica gel using 3ū75 mL of ethyl acetate followed by 5:1 ethyl acetate-ethanol. (+)-FTC (2) was isolated as a white solid (109 mg; 37% based on starting butyrate). HPLC analysis of the (+)-FTC as its 5'-O-acetate derivative showed 97.4% e.e.; a!O(20,D) +113.4░ (c=2.53; absolute ethanol)
A similar reaction was performed using 0.12 mmol (37 mg) of the 5'-O-butyrate of FTC and 7 mg of PS-800 in 4.0 mL of 4:1 pH 8 buffer:CH3 CN. The reaction was considerably slower than that with PLE-A and required 74 hours for 59% conversion. The recovered butyrate (11.4 mg; 31% of the initial amount) was found to be 94% e.e. by HPLC.
In an alternative embodiment, cytidine-deoxycytidine deaminase is used to resolve racemic mixtures of 2-hydroxymethyl- 5-(cytosin-1-yl)-1,3-oxathiolane and its derivatives, including 2-hydroxymethyl-5-(5-fluoro-cytosin-1-yl)-1,3-oxathiolane. The enzyme catalyses the deamination of the cytosine moiety to a uridine. It has been discovered that one of the enantiomers of 1,3-oxathiolane nucleosides is a preferred substrate for cytidine-deoxycytidine deaminase. The enantiomer that is not converted to a uridine (and therefore is still basic) is extracted from the solution with an acidic solution. Care should be taken to avoid strong acidic solutions (pH below 3.0), that may cleave the oxathiolane ring.
Cytidine-deoxycytidine deaminase can be isolated from rat liver or human liver, or expressed from recombinant sequences in procaryotic system such as in E. coli.
The method of resolution of cytidine nucleoside enantiomers using cytidine-deoxycytidine deaminase can be used as the sole method of resolution or can be used in combination with other methods of resolution, including resolution by enzymatic hydrolysis of 5'-O-nucleoside esters as described above.
The process described above for resolving racemic mixtures of nucleoside enantiomers can be combined with other classical methods of enantiomeric resolution to increase the optical purity of the final product.
Classical methods of resolution include a variety of physical and chemical techniques. Often the simplest and most efficient technique is recrystallization, based on the principle that racemates are often more soluble than the corresponding individual enantiomers. Recrystallization can be performed at any stage, including on the acylated compounds and or the final enantiomeric product. If successful, this simple approach represents a method of choice.
When recrystallization fails to provide material of acceptable optical purity, other methods can be evaluated. If the nucleoside is basic (for example, a cytidine) one can use chiral acids that form diastereomeric mixtures that may possess significantly different solubility properties. Nonlimiting examples of chiral acids include malic acid, mandelic acid, dibenzoyl tartaric acid, 3-bromocamphor-8-sulfonic acid, 10-camphorsulfonic acid, and di-p-toluoyltartaric acid. Similarly, acylation of the free hydroxyl group with a chiral acid derivative also results in the formation of diastereomeric mixtures whose physical properties may differ sufficiently to permit separation.
Small amounts of enantiomerically enriched nucleosides can be obtained or purified by passing the racemic mixture through an HPLC column that has been designed for chiral separations, including cyclodextrin bonded columns marketed by Rainin Corporation.
The resolutions of the C4'-enantiomers of (▒)-BCH-189 and (▒)-FTC were performed using a chiral cyclodextrin bonded (cyclobond AC-I) column obtained from Rainin Corporation (Woburn, MA). The conditions were as follows: Isocratic 0.5% methanol in water; flow rate 1 ml/min., UV detection at 262 nm. HPLC grade methanol was obtained from J. T. Baker (Phillipsburg, N.J.). The racemic mixtures were injected and fractions were collected. Fractions containing each of the enantiomers were pooled, frozen, and then lyophilized. The compounds were characterized by UV spectroscopy and by their retention times on HPLC. In general, the (-)-enantiomers have lower retention times than the (+)-enantiomers (see J. Liquid Chromatoqraphy 7:353 -376, 1984). The concentrations of the compounds were determined by UV spectroscopy, using a stock solution of known concentration (15 μM) prepared in water for biological evaluation. The retention times for the separated enantiomers are provided in Table 2.
TABLE 2______________________________________Retention Times of Enantiomers of BCH-189 and FTCCompound R, (min)______________________________________(-)-BCH-189 9.0(+)-BCH-189 10.0(-)-FTC 8.3(+)-FTC 8.7______________________________________
II. Antiviral Activity of 2-Hydroxymethy1-5-(5-Fluorocytosin-1-yl)-1,3-oxathiolane ("FTC")
It is often desirable to screen a number of racemic mixtures of nucleosides as a preliminary step to determine which warrant further resolution into enantiomerically enriched components and evaluation of antiviral activity. The ability of nucleosides to inhibit HIV can be measured by various experimental techniques. The technique used herein, and described in detail below, measures the inhibition of viral replication in phytohemagglutinin (PHA) stimulated human peripheral blood mononuclear (PBM) cells infected with HIV-1 (strain LAV). The amount of virus produced is determined by measuring the virus- coded reverse transcriptase enzyme. The amount of enzyme produced is proportional to the amount of virus produced. Table 4 provides the EC50 values (concentration of nucleoside that inhibits the replication of the virus by 50% in PBM cells) and IC50 values (concentration of nucleoside that inhibits 50% of the growth of mitogen-stimulated uninfected human PBM cells) of a number of (▒)-1,3-oxathiolane and nucleosides.
A. Three-day-old phytohemagglutinin-stimulated PBM cells (106 cells/ml) from hepatitis B and HIV-1 seronegative healthy donors were infected with HIV-1 (strain LAV) at a concentration of about 100 times the 50% tissue culture infectious dose (TICD 50) per ml and cultured in the presence and absence of various concentrations of antiviral compounds.
B. Approximately one hour after infection, the medium, with the compound to be tested (2 times the final concentration in medium) or without compound, was added to the flasks (5 ml; final volume 10 ml). AZT was used as a positive control.
C. The cells were exposed to the virus (about 2ū105 dpm/ml, as determined by reverse transcriptase assay) and then placed in a CO2 incubator. HIV-1 (strain LAV) was obtained from the Center for Disease Control, Atlanta, Ga. The methods used for culturing the PBM cells, harvesting the virus and determining the reverse transcriptase activity were those described by McDougal et al. (J. Immun. Meth. 76, 171-183, 1985) and Spira et al. (J. Clin. Meth. 25, 97-99, 1987), excepts that fungizone was not included in the medium (see Schinazi, et al., Antimicrob. Agents Chemother. 32, 1784-1787 (1988); Id., 34:1061-1067 (1990)) .
D. On day 6, the cells and supernatant were transferred to a 15 ml tube and centrifuged at about 900 g for 10 minutes. Five ml of supernatant were removed and the virus was concentrated by centrifugation at 40,000 rpm for 30 minutes (Beckman 70.1 Ti rotor). The solubilized virus pellet was processed for determination of the levels of reverse transcriptase. Results are expressed in dpm/ml of sampled supernatant. Virus from smaller volumes of supernatant (1 ml) can also be concentrated by centrifugation prior to solubilization and determination of reverse transcriptase levels.
The median effective (EC50) concentration was determined by the median effect method (Antimicrob. Agents Chemother. 30, 491-498 (1986). Briefly, the percent inhibition of virus, as determined from measurements of reverse transcriptase, is plotted versus the micromolar concentration of compound. The EC50 is the concentration of compound at which there is a 50% inhibition of viral growth.
E. Mitogen stimulated uninfected human PBM cells (3.8ū105 cells/ml) were cultured in the presence and absence of drug under similar conditions as those used for the antiviral assay described above. The cells were counted after 6 days using a hemacytometer and the trypan blue exclusion method, as described by Schinazi et al., Antimicrobial Agents and Chemotherapy, 22(3), 499 (1982). The IC50 is the concentration of compound which inhibits 50% of normal cell growth.
TABLE 3______________________________________EC50 and IC50 of Various Analogues of 1,3-OxathiolaneNucleosides in human PBM cells. Antiviral CytotoxicityCode X or Y R EC50, μM IC50, μM______________________________________DLS-009 X = O H >100 >100DLS-010 X = O Me 64.4 >100DLS-027 X = O F >100 >100DLS-028 X = O Cl 60.8 >100DLS-044 X = O Br >100 >100DLS-029 X = O I >100 >100DLS-020 Y = NH2 H 0.02 >100DLS-011 Y = NH2 Me >10 >100DLS-022 Y = NH2 F 0.011 >100DLS-023 Y = NH2 Cl 38.7 >100DLS-021 Y = NH2 Br 77.4 >100DLS-026 Y = NH2 I 0.72 >100DLS-058(-) Y = NH2 F 0.0077 >100DLS-059(+) Y = NH2 F 0.84 >100DLS-053 Y = NH2 CF3 60.7 >100______________________________________
As indicated in Table 3, in general, the substituted cytosine 1,3-oxathiolane nucleosides are more active than the corresponding uracil nucleosides. One of the compounds, (▒)-FTC, (referred to as "DLS-022", compound 8) not only exhibited exceptional activity (approximately 10 nM in PBM cells), but also quite low toxicity (>100 μM in PBM, Veto and CEM cells). Ths activity compares quite favorably with 2',3'-dideoxyadenosine (DDA, EC50 =0.91 μM), 3'-azido2',3'-dideoxyuridine (AZDU, EC50 =0.18-0.46 μM), 3'-dideoxythymidine (DDT, EC50 =0.17 μM), and dideoxycytidine (DDC, EC50 =0.011 μM).
The IC50 of (▒)-FTC was measured as over 100 μM, indicating that the compound was not toxic in uninfected PBM cells evaluated up to 100 μM.
The enantiomers of FTC were isolated by the method of Example 3, and the antiviral activity evaluated by the method of Example 4. The results are provided in Table 4, and illustrated in FIG. 4.
TABLE 4______________________________________Antiviral Activity of the (+) and (-) Enantiomers of FTC % Inhibition EC50 :μMTreatment Concn., μM DPM/ml (Corrected)______________________________________FTC (▒) 0.0001 73,755 26.6 0.018 0.005 83,005 16.3 0.01 60,465 41.3 0.05 34,120 70.4 0.1 14,160 92.4 0.5 18,095 88.1 1 7,555 99.7 5 7,940 99.3 10 5,810 101.7FTC (-) 0.001 76,275 23.8 0.02 0.005 58,590 43.3 0.01 75,350 24.8 0.05 28,890 76.2 0.1 13,175 93.5 0.5 9,485 97.6FTC (+) 0.001 94,340 3.8 0.28 0.005 107,430 -10.6 0.01 99,465 -1.8 0.05 87,120 11.8 0.1 86,340 12.7 0.5 33,225 71.4______________________________________
As indicated in Table 4, the (-)-enantiomer of FTC is approximately one order of magnitude more potent than the (+)-FTC enantiomer, and has approximately the same anti-HIV activity as the racemic mixture. Neither the enantiomers nor the racemic mixture is toxic up to 100 μM as measured by Trypan Blue exclusion in human PBM cels.
The enantiomers of (▒)-FTC were also resolved by the method of Example 2, and the antiviral activity evaluated by the method of Example 4. The results are illustrated in FIG. 5. As indicated in FIG. 5, the EC50 of the racemic mixture of FTC was measured at 0.017 μM, the ECH50 of (-)-FTC at 0.0077 μM, and the EC50 of (+)-FTC at 0.84 μM.
The differences in EC50 as measured in Examples 5 and 6 may be due to a number of factors, including differences in donor PBM cells, the inherent error of the anti-HIV screening procedure (estimated at approximately 10%), and differences in the measurement of concentration of the nucleosides as resolved in the methods of Examples 2 and 3. In the method of Example 2, the FTC enantiomers were isolated as solids and weighed to prepare the testing solution. In the method of Example 3, the concentration of the FTC enantiomers was estimated from UV absorption measurements.
The data indicates that the (+) enantiomer is significantly less potent than the (-) entantiomer or the racemic mixture.
The enantiomers of BCH-189 were isolated by the method of Example 3, and the antiviral activity evaluated by the method of Example 4. The results are provided in Table 5, and illustrated in FIG. 6.
TABLE 5______________________________________Antiviral Activity of the (+) and (-) Enantiomers of BCH-189 % Inhibition EC50 :μMTreatment Concn., μM DPM/ml (Corrected)______________________________________Blanks mean 762HIV Std. 158,705Uninfected mean 7,320Control ▒SD 4,520Infected mean 97,795Control ▒SD 6,790BCH-189 (▒) 0.001 65,170 36.1 0.081 0.005 62,595 38.9 0.01 70,875 29.8 0.05 77,650 22.3 0.1 33,165 71.4 0.5 10,765 96.2 1 7,745 99.5 5 6,800 100.6 10 4,470 103.2BCH-189 (-) 0.001 76,400 23.6 0.016 0.005 66,875 34.2 0.01 54,170 48.2 0.05 57,615 44.4 0.1 34,705 69.7 0.5 15,250 91.2BCH-189 (+) 0.00085 71,795 28.7 0.23 0.00425 99,710 -2.1 0.0085 68,355 32.5 0.0415 82,845 16.5 0.0825 65,100 36.1 0.412 43,260 60.3______________________________________
As indicated in Table 6, the (-)-enantiomer of BCH-189 is approximately one order of magnitude more potent than the (+)-FTC enantiomer, and has approximately the same anti-HIV activity as the racemic mixture. Neither enantiomer exhibited any toxicity in a concentration up to 100 μM as measured by Trypan Blue exclusion in human PBM cells.
Studies were undertaken using radiolabeled agent in order to follow th eintracellular profiles of the parent drug and metabolites detected within the cell. All studies were conducted in duplicate. Human peripheral blood mononuclear cells (PBM cells) are suspended in RPMI 1640 medium containing 10% getal calf serum and antibiotics (2ū106 cells/ml), 10 ml per timepoint) and incubated with addition of 10 μM FTC (specific ctivity about 700 dpm/pmol). Cells are exposed to the drug for 2, 6, 12 and 24 hours. At these timepoints, the medium is removed and the cells are washed two times with cold Hank's balanced salt solution. Extraction is performed with addition of 0.2 ml of 60% cold methanol/water and stored overnight at -70 ░ C. The following morning, the suspensions are centrifuged and extractions are repeated two times for 0.5 hours at -70 ░ C. The total supernatants (0.6 ml) are lyophilized to dryness. The residues are resuspended in 250 μl of water and aliquots comprising between 50 and 100 μl are analyzed by HPLC. Quantitation of intracellular parent drug and metabolic derivatives are conducted by HPLC. Because of the potential acid lability of some compounds, a buffer system close to physiological pH is used for the separation of the metabolites.
FIG. 7 is a graph of the uptake of tritiated (▒)-FTC in human PBM cells (average of two determinations) in time (hours) versus pmol/106 cells. The uptake studies indicate that radiolabeled FTC is readily taken up in human lymphocytes, that produce very large amounts of 5'-triphosphate.
The same procedure was used with human liver cells as with PBM cells to determine uptake of FTC.
The (▒)-FTC is taken up by hepG2 cells in large amounts, and that these human liver cells metabolize a large percentage of the (▒)-FTC to (▒)-FTC triphosphate.
These data in conjunction with other data indicate that (▒)-FTC, as well as its (-) and (+) enantiomers, are effective as antiviral agents against HBV (hepatitis B virus).
Studies were performed using radiolabeled FTC in order to follow the intracellular profiles of the parent drug and metabolites detected within the cell after removal of drug at different times after pulsing for 24 hours, the time needed for high levels of triphosphates to accumulate. Studies are conducted in duplicate. Uninfected cells (2 ū106 ml) are suspended in the appropriate medium supplemented with serum (10 ml per timepoint) and incubated at 37░ C. in a 5% CO2 incubator. Radiolabeled FTC concentration is 10 μM. After pulsing the cells with the labeled compound for the desired time, the cells are thoroughly washed and then replenished with fresh medium without the antiviral drugs (0 hr). At 0, 2, 4, 6, 12, 24, and 48 hours (second incubation time), the cells are removed, and immediately extracted with 60% cold methanol/water. The extract is obtained by centrifugation and removal of the cell pellet. The extracts are lyophilized and then stored at -70░ C. Prior to analysis, the material is resuspended in 250 τl of HPLC buffer and immediately analyzed. Quantitation of intracellular parent drug and metabolic derivatives are conducted by HPLC, as follows.
Either a Micromeritics or Hewlett-Packard model 1090 PHLC system is used with an anion exchange Partisil 10 SAX column (Whatman., Inc.), at a flow rate of 1 ml/min, 1 kpsi pressure, using uv detection at 262 nm.
The mobile phase consists of:
a. deionized water
b. 2 mM NaH2 PO4 /16 mM NaOAc (pH=6.6)
c. 15 mM NaH2 PO4 /120.2 mM NaOAc (pH=6.6)
d. 100 mM NaH2 PO4 /800 mM NaOAc (pH=6.6)
Separation method: isocratic for 5 min with A, followed by a 15 min linear gradient to 100% B, followed by a 20 min linear gradient to 100% C, followed by 10 min linear gradient to 100% D, followed by 30 min isocratic with 100% D.
______________________________________Retention times (minutes) in human cells: Mono-Compound Unchanged phosphate Diphosphate Triphosphate______________________________________DLS-022 5.0 39.0 55.0 68.0BCH-189 3.5 40.0 55.0 69.0______________________________________
FIG. 8 is a graph of the egress of radiolabeled (▒)-FTC from human PBM cells, measured in hours after drug removal versus concentration (pmol/106 cells). As indicated in the Figure, FTC-triphosphate has an intracellular half-life of approximately 12 hours and can be easily detected intracellularly at concentrations of 1-5 μM 48 hours after the removal of the extracellular drug, which is well above the EC50 for the compound. Further, the affinity (K') for (▒)-FTC triphosphate against HIV RT is 0.2 μM, below the 48 hour concentration level.
III. Preparation of Pharmaceutical Compositions.
Humans suffering from diseases caused by HIV infection can be treated by administering to the patient an effective amount of (▒)-FTC, or its (-) or (+) enantiomer or a pharmaceutically acceptable salt thereof in the presence of a pharmaceutically acceptable carrier or diluent. The active materials can be administered by any appropriate route, for example, orally, parenterally, intravenously, intradermally, subcutaneously, or topically, in liquid or solid form.
Pharmaceutically acceptable salts are known to those in the art and include those derived from pharmaceutically acceptable inorganic and organic acids and bases. Examples of suitable acids include hydrochloric, hydrobromic, sulfuric, nitric, perchloric, fumaric, maleic, phosphoric, glycollic, lactic, salicyclic, succinic, toluene-p-sulfonic, tartaric, acetic, citric, methanesulfonic, formic, benzoic, malonic, naphthalene-2-sulfonic and benzenesulfonic acids. Salts derived from appropriate bases include alkali metal (e.g., sodium), alkaline earth metal (e.g., magnesium), ammonium and quaternary amine.
The active compound is included in the pharmaceutically acceptable carrier or diluent in an amount sufficient to deliver to a patient a therapeutically eoffective amount of compound to inhibit viral replication in vivo, especially HIV and HBV replication, without causing serious toxic effects in the patient treated. By "HIV inhibitory amount" is meant an amount of active ingredient sufficient to exert an HIV inhibitory effect as measured by, for example, an assay such as the ones described herein.
A preferred dose of (-) or (▒)-FTC will be in the range from about 1 to 20 mg/kg of bodyweight per day, more generally 0.1 to about 100 mg per kilogram body weight of the recipient per day.
The compound is conveniently administered in unit dosage form: for example containing 7 to 7000 mg, preferably 70 to 1400 mg of active ingredient per unit dosage form.
Ideally the active ingredient should be administered to achieve peak plasma concentrations of the active compound of from about 0.2 to 70 μM, preferably about 1.0 to 10 μM. This may be achieved, for example, by the intravenous injection of a 0.1 to 5% solution of the active ingredient, optionally in saline, or administered as a bolus of the active ingredient.
The concentration of active compound in the drug composition will depend on absorption, inactivation, and excretion rates of the drug as well as other factors known to those of skill in the art. It is to be noted that dosage values will also vary with the severity of the condition to be alleviated. It is to be further understood that for any particular subject, specific dosage regimens should be adjusted over time according to the individual need and the professional judgment of the person administering or supervising the administration of the compositions, and that the concentration ranges set forth herein are exemplary only and are not intended to limit the scope or practice of the claimed composition. The active ingredient may be administered at once, or may be divided into a number of smaller doses to be administered at varying intervals of time. A preferred mode of administration of the active compound is oral. Oral compositions will generally include an inert diluent or an edible carrier. They may be enclosed in gelatin capsules or compressed into tablets. For the purpose of oral therapeutic administration, the active compound can be incorporated with excipients and used in the form of tablets, troches, or capsules. Pharmaceutically compatible binding agents, and/or adjuvant materials can be included as part of the composition.
The tablets, pills, capsules, troches and the like can contain any of the following ingredients, or compounds of a similar nature: a binder such as microcrystalline cellulose, gum tragacanth or gelatin; an excipient such as starch or lactose, a disintegrating agent such as alginic acid, Primogel, or corn starch; a lubricant such as magnesium stearate or Sterotes; a glidant such as colloidal silicon dioxide; a sweetening agent such as sucrose or saccharin; or a flavoring agent such as peppermint, methyl salicylate, or orange flavoring. When the dosage unit form is a capsule, it can contain, in addition to material of the above type, a liquid carrier such as a fatty oil. In addition, dosage unit forms can contain various other materials which modify the physical form of the dosage unit, for example, coatings of sugar, shellac, or other enteric agents. (▒)-FTC, or its (-) or (+)-enantiomer or pharmaceutically acceptable salts thereof can be administered as a component of an elixir, suspension, syrup, wafer, chewing gum or the like. A syrup may contain, in addition to the active compounds, sucrose as a sweetening agent and certain preservatives, dyes and colorings and flavors.
(▒)-FTC, or its (-) or (+)-enantiomers, or pharmaceutically acceptable salts thereof can also be mixed with other active materials that do not impair the desired action, or with materials that supplement the desired action, such as antibiotics, antifungals, antiinflammatories, or other antivirals, including other nucleoside anti-HIV compounds.
Solutions or suspensions used for parenteral, intradermal, subcutaneous, or topical application can include the following components: a sterile diluent such as water for injection, saline solution, fixed oils, polyethylene glycols, glycerine, propylene glycol or other synthetic solvents; antibacterial agents such as benzyl alcohol or methyl parabens; antioxidants such as ascorbic acid or sodium bisulfite; chelating agents such as ethylenediaminetetraacetic acid; buffers such as acetates, citrates or phosphates and agents for the adjustment of tonicity such as sodium chloride or dextrose. The parental preparation can be enclosed in ampoules, disposable syringes or multiple dose vials made of glass or plastic.
The pharmaceutical composition can also include antifungal agents, chemotherapeutic agents, and other antiviral agents such as interferon, including α, β, and gamma interferon.
If administered intravenously, preferred carriers are physiological saline or phosphate buffered saline (PBS).
In a preferred embodiment, the active compounds are prepared with carriers that will protect the compound against rapid elimination from the body, such as a controlled release formulation, including implants and microencapsulated delivery systems. Biodegradable, biocompatible polymers can be used, such as ethylene vinyl acetate, polyanhydrides, polyglycolic acid, collagen, polyorthoesters, and polylactic acid. Methods for preparation of such formulations will be apparent to those skilled in the art. The materials can also be obtained commercially from Alza Corporation and Nova Pharmaceuticals, Inc.
Liposomal suspensions (including liposomes targeted to infected cells with monoclonal antibodies to viral antigens) are also preferred as pharmaceutically acceptable carriers. These may be prepared according to the methods known to those skilled in the art, for example, as described in U.S. Pat. No. 4,522,811 (which is incorporated herein by reference in its entirety). For example, liposome formulations may be prepared by dissolving appropriate lipid(s) (such as stearoyl phosphatidyl ethanolamine, stearoyl phosphatidyl choline, arachadoyl phosphatidyl choline, and cholesterol) in an inorganic solvent that is then evaporated, leaving behind a thin film of dried lipid on the surface of the container. An aqueous solution of the active compound or its monphosphate, diphosphate, and/or triphosphate derivatives are then introduced into the container. The container is then swirled by hand to free lipid material from the sides of the container and to disperse lipid aggregates, thereby forming the liposomal suspension.
IV. Preparation of Phosphate Derivatives of FTC
Mono, di, and triphosphate derivative of FTC can be prepared as described below.
The monophosphate can be prepared according to the procedure of Imai et al., J. Org. Chem., 34(6), 1547-1550 (June 1969). For example, about 100 mg of FTC and about 280 μl of phosphoryl chloride are reacted with stirring in about 8 ml of dry ethyl acetate at about 0░ C. for about four hours. The reaction is quenched with ice. The aqueous phase is purified on an activated charcoal column, eluting with 5% ammonium hydroxide in a 1:1 mixture of ethanol and water. Evaporation of the eluant gives ammonium FTC-5'-monophosphate.
The diphosphate can be prepared according to the procedure of Davisson et al., J. Org. Chem., 52(9), 1794-1801 (1987). FTC diphosphate can be prepared from the corresponding tosylate, that can be prepared, for example, by reacting the nucleoside with tosyl chloride in pyridine at room temperature for about 24 hours, working up the product in the usual manner (e.g., by washing, drying, and crystallizing it).
The triphosphate can be prepared according to the procedure of Hoard et al., J. Am. Chem. Soc., 87(8), 1785-1788 (1965). For FTC is activated (by making a imidazolide, according to methods known to those skilled in the art) and treating with tributyl ammonium pyrophosphate in DMF. The reaction gives primarily the triphosphate of the nucleoside, with some unreacted monophosphate and some diphosphate. Purification by anion exchange chromatography of a DEAE column is followed by isolation of the triphosphate, e.g., as the tetrasodium salt.
This invention has been described with reference to its preferred embodiments. Variations and modifications of the invention, a method of resolution and antiviral activity of nucleoside enantiomers, will be obvious to those skilled in the art from the foregoing detailed description of the invention. It is intended that all of these variations and modifications be included within the scope of the appended claims.
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Idaho soldier freed from captivity in Afghanistan
The circumstances surrounding Bergdahl's capture remain something of a mystery. There has been some speculation that he willingly walked away from his unit, raising the question of whether he could be charged with being absent without leave or desertion. A senior U.S. official told The Associated Press on Saturday that the Army would make the decision on any charges but that the feeling at the moment was that Bergdahl had suffered enough in his ordeal. The official was not authorized to discuss the matter by name and requested anonymity.
In 2012, Rolling Stone magazine quoted emails Bergdahl is said to have sent to his parents that suggest he was disillusioned with America's mission in Afghanistan, had lost faith in the U.S. Army's mission there and was considering desertion. Bergdahl told his parents he was "ashamed to even be American."
The Associated Press could not independently authenticate the emails.
Were Bergdahl to be charged with desertion, the maximum penalty he would face is five years in prison and a dishonorable discharge, if it's proven that he deserted with the intent to avoid hazardous duty or to shirk important service. A case of AWOL, ended by the U.S. apprehending him, would not require proof that he intended to remain away permanently. The maximum punishment for that would be a dishonorable discharge and 18 months' confinement, according to military justice experts.
The U.S. has long been seeking Bergdahl's release, but there was renewed interest in his release as Obama finalized plans to pull nearly all American forces out of Afghanistan by the end of 2016.
Officials said the Taliban signaled to the U.S. in November that they were ready to start new talks on the issue of detainees. After the U.S. received proof that Bergdahl was still alive, indirect talks began, with Qatar sending messages back and forth between the two parties.
The talks intensified about a week ago, officials said, resulting in Bergdahl's release and the transfer of the Afghan detainees.
The senior U.S. official said those U.S. officials involved in the swap decided it could further the effort to reach reconciliation with the Taliban, which they see as a key to achieving a higher level of security in Afghanistan. They acknowledged in their discussions the question of emboldening other insurgent groups to take troops or other Americans prisoner to secure the release of other prisoners, according to the official. The military believed that Bergdahl's status as a prisoner of war obliged doing whatever possible to obtain his release, the official said.
The five Guantanamo detainees departed the base on a U.S. military aircraft Saturday afternoon. Under the conditions of their release, the detainees will be banned from traveling outside of Qatar for at least one year.
Obama and the emir of Qatar spoke last week about the conditions of the release, which have been codified in a memorandum of understanding between the two countries, officials said.
The administration is legally required to notify Congress in advance about plans to release Guantanamo detainees. An administration official said lawmakers were notified only after U.S. officials knew they had Bergdahl, but before the transfers took place.
Two Republican lawmakers said Obama violated U.S. laws when he approved the exchange. Rep. Howard "Buck" McKeon of California and Sen. James Inhofe of Oklahoma said the law required Obama to notify Congress 30 days before any transfer of terrorists from the U.S. detention facility at Guantanamo Bay, Cuba. In response, the White House said that officials considered what they called "unique and exigent circumstances" and decided to go ahead with the transfer in spite of the legal requirement.
The detainees are among the most senior Afghans still held at the prison. They are:
—Abdul Haq Wasiq, who served as the Taliban deputy minister of intelligence
—Mullah Norullah Nori, a senior Taliban commander in the northern city of Mazar-e-Sharif when the Taliban fought U.S. forces in late 2001
—Khairullah Khairkhwa, who served in various Taliban positions including interior minister and had direct ties to Mullah Omar and Osama bin Laden
—Mohammed Nabi, who served as chief of security for the Taliban in Qalat, Afghanistan, and later worked as a radio operator for the Taliban's communications office in Kabul
—Mohammad Fazl, whom Human Rights Watch says could be prosecuted for war crimes for presiding over the mass killing of Shiite Muslims in Afghanistan in 2000 and 2001 as the Taliban sought to consolidate their control over the country.

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Archive September 2009
NZ's largest science blog network goes live Sep 30No Comments
You will notice a rash of new science blogs based in New Zealand. Maybe you have already.
This press release explains it all.
The Science Media Centre today launches a major new science communication effort with the arrival of Sciblogs, a network of science blogs covering everything from clinical health to climate change.
Sciblogs debuts with 26 bloggers, including scientists from universities, Crown Research Institutes and private research organisations the length of the country. It will be the largest online hub for science-related content relevant to New Zealanders and act as a forum of discussion on the important issues facing society.
Syndicating their work through Sciblogs are well-established bloggers like Open Parachute's Dr Ken Perrott, MacDoctor's Dr Jim McVeagh, Hot Topic's Gareth Renowden and Dr Alison Campbell, the founder of BioBlog.
Newcomers joining the line-up include University of Otago marine ecologist and 2008 MacDiarmid Young Scientist of the Year Dr Rebecca McLeod, Victoria University climate scientist Dr Andy Reisinger and astrophysicist Dr Shaun Hendy, who is deputy director of the MacDiarmid Institute for Advanced Materials and Nanotechnology.
A full list of our bloggers can be found here.
"In business and politics, health and agriculture, science features prominently in all aspects of our lives these days," says Peter Griffin, manager of the Science Media Centre.
"But often there's little decent discussion of the science itself. New Zealand has some great science bloggers, but they don't get the recognition they deserve because they are scattered across the web."
Sciblogs brings the best science bloggers together in one place to form the largest local online community dedicated to science. Sciblog contributors have full editorial and creative control of their blogs and new Scibloggers will be added over time.
The Sciblogs platform, built by Auckland web developer Marker Studio, is the first major implementation of the opensource WordPress MU (multiple user) blogging system in New Zealand.
"The system is totally scalable so we can add new bloggers in a matter of minutes," says Griffin.
Ultimately the goal is to start ongoing conversations on the big issues, everything from genetic modification and climate science to health care and our use of new technology.
"It's a bit of an experiment for New Zealand where the blogosphere is dominated by political pundits and where the conversation can often turn nasty," says Griffin.
"But we think there's an appetite for intelligent comment on scientific issues and Sciblogs will be central to that."
And here's a list of the current blogs – do go and have a browse:
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- NZ science bloggers – new opportunity (openparachute.wordpress.com)
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Sustainability and ethics Sep 30No Comments
Review: Strong sustainability for New Zealand : principles and scenarios, by Sustainable Aotearoa New Zealand Inc. (SANZ).
NZ Publisher: Nakedize Limited
Binding: paperback
Pages: 52
PDF available for download here. Briefing (with audio) here.
I think almost all of us will admit, in our more honest moments, that we can't go on as we are. 'Business as usual' is just not a long-term alternative — ethically as well as economically.
Most attention these days focuses on climate change. But, this is just part of a wider problem resulting from our influence on the environment. Climate change, the undermining of biological diversity, the problems of waste production and resource depletion.
Sustainable economics seems the obvious way of confronting these problems. But what this is, and how to do it, can be controversial.
New ethics for sustainability
Sustainable Aotearoa NZ (SANZ) has launched this paper with to promote discussion of our sustainable development. Inevitably much of the content will be considered controversial. It's meant to be. SANZ hopes it will be used as a starting point to further discussion. In particular it is promoting the idea that sustainable development is not just a matter of economics. It will need a whole new way of thinking about economics and personal responsibility and lifestyles. It needs a new ethics.
And this is the main message of this paper. Not surprising, because it is one of the main points promoted by SANZ. Another is its advocacy of 'strong sustainability.'
Besides an introduction the paper is divided into a part on principles of strong sustainability, a scenario for transition to strong sustainability and (the largest section) a scenario of a strongly sustainable NZ.
It is this last section which provokes. It restricts itself to just a 'strong sustainability' scenario with matching wide acceptance of the needed ethical attitudes by the population. It doesn't consider 'weaker' alternatives because the writers consider them to be ineffective, leading only to disaster.
The paper envisages a society where we live in harmony with nature. Where many of the environmentally threatening aspects of the economy no longer exist. Employment though is no longer seen to be a problem as many roles will be more labour-intensive. Working hours will be shorter and there will be more leisure time. Communities will be structured to require less travel.
Although global climate change will have less direct affect in New Zealand than elsewhere we will have to adjust to huge changes in global trade and migration.
Predicting the future?
It's always difficult to predict the future. One just has to consider the unprecedented, and unpredicted, details of the technology change we have all experienced over the last 30 years. So many readers will disagree with details of this scenario. For example, the widespread change to 'organic farming' methods will probably appeal to some but may be less sensible than forms of sustainable factory production of food.
Similarly this scenario will draw political and ideologically motivated responses. Many people will accept as inevitable the departure from 'business as usual' model of unrestricted. But they may not like the seeming inevitability of a more socialist society. And of course there will be those who see this scenario as an expression of a green 'religious' approach.
However, ideological and political conflict always accompany social change. These conflicts are unavoidable because sustainability will require big changes. For the sake of our future I hope that a broad support for sustainability can develop despite this conflict.
New approaches to sustainable approaches to current economic, social and political questions will be needed in the transition to sustainability. One weakness of this paper is that current questions get little consideration. Political and social involvement of those promoting sustainability will be needed to ensure such voices are heard.
The paper itself is presented in a format more familiar to glossy corporate reports. Reports meant to impress but often not read. Such glossy productions are not amenable to practical use — try annotating documents like this. And SANZ is probably effective in its activity directed at commerce and academia. But dissemination of their arguments would be helped by on-line forums and blogs suitable for involvement of individuals in discussion of their policies.
Nevertheless, this paper will contribute to the wider political and social discussion. It, and the ideas presented, need to get into mainstream forums and publications. This will help to promote the discussion we need to have.
Our future depends on it.
See also: SANZ – Sustainable Development in New Zealand.
NZ blogs sitemeter ranking — September '09 Sep 28No Comments
Currently I have found 113 NZ blogs with publicly accessible sitemeter stats. This is about 20% of the total on my database.
The blogs are listed in the table below, together with daily visits and page view numbers averaged over the previous 7 days. The data was that given by by the NZ blog ranking tool on September 26th.
This can be seen at NZ blogs average daily visits
Have a look at that tool. Its a way of comparing your own blog's performance from day to day - a roller coaster ride which can be quite exciting!
Just be aware that it relies on spreadsheets ad Googles Docs and data is not always available. If the data is missing or incomplete, wait a while and reload the page. it will eventually show up.
Meanwhile I am still keen to hear of any other blogs with publicly available sitemeter stats that I have missed. Contact me if you know of any.
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The naked emperor Sep 27No Comments
It's interesting how a simple word, a name or a book title can cause twitching in the knee area. I predicted we would see a rash of this syndrome with the publication of Richard Dawkins new book The Greatest Show on Earth: The Evidence for Evolution. We are going to see more of the syndrome as the book reviews and other publicity appear in our newspapers and magazines.
One of the most common symptoms of the knee jerking goes right back the Terry Eagleton's early review of Dawkins' previous book, The God Delusion. That is the charge that Dawkins had no right to produce that book because he is not a religious philosopher or theologian (see Do you believe in a god?).
I think this symptom also indicates laziness on the part of the sufferer. The delusion that a review of titles, or ones own prejudices, will suffice rather than a proper review of the book. The syndrome seems more common in individuals also suffering from other afflictions involving mystical beliefs, or conviction of the existence of gods who look out for them.
Even though Dawkins' new book does not deal directly with these mystical beliefs those knees are still jerking – as readers will note from recent comments here (see Dawkins bashing season upon us?). I thought it would be interesting to reintroduce something written by PZ Myers, author of the science blog Pharyngula, three years ago - The Courtier's Reply. Many of you will have read it before but its a great piece of writing and deserves to be more widely known. So I repeat it below:
"I have considered the impudent accusations of Mr Dawkins with exasperation at his lack of serious scholarship. He has apparently not read the detailed discourses of Count Roderigo of Seville on the exquisite and exotic leathers of the Emperor's boots, nor does he give a moment's consideration to Bellini's masterwork, On the Luminescence of the Emperor's Feathered Hat. We have entire schools dedicated to writing learned treatises on the beauty of the Emperor's raiment, and every major newspaper runs a section dedicated to imperial fashion; Dawkins cavalierly dismisses them all. He even laughs at the highly popular and most persuasive arguments of his fellow countryman, Lord D. T. Mawkscribbler, who famously pointed out that the Emperor would not wear common cotton, nor uncomfortable polyester, but must, I say must, wear undergarments of the finest silk.
Dawkins arrogantly ignores all these deep philosophical ponderings to crudely accuse the Emperor of nudity.
Personally, I suspect that perhaps the Emperor might not be fully clothed – how else to explain the apparent sloth of the staff at the palace laundry – but, well, everyone else does seem to go on about his clothes, and this Dawkins fellow is such a rude upstart who lacks the wit of my elegant circumlocutions, that, while unable to deal with the substance of his accusations, I should at least chide him for his very bad form.
Until Dawkins has trained in the shops of Paris and Milan, until he has learned to tell the difference between a ruffled flounce and a puffy pantaloon, we should all pretend he has not spoken out against the Emperor's taste. His training in biology may give him the ability to recognize dangling genitalia when he sees it, but it has not taught him the proper appreciation of Imaginary Fabrics."
By the way. Myers is currently writing a book. I imagine we will see it some time next year. He certainly has great literary skill so I imagine it will be a best seller.
I look forward to it.
From the keyboards of scientists… Sep 25No Comments
This from the Science Media Centre NZ:
Are scientists blogging in New Zealand? They sure are – but it is relatively tricky to find them.
That will change on September 30 when the Science Media Centre launches Sciblogs.co.nz, a blogging network bringing together some of the best science blogs in the country and launching a good few new ones.
Sciblogs will kick off with 25 bloggers from universities, CRIs and private research institutions covering everything from healthcare to climate change. Our bloggers know their stuff – they have two dozen PhDs between them and they aren't afraid to tackle the big science-related issues affecting society.
Sciblogs will become an online hub for discussion of scientific topics and will grow to include other bloggers. If you would like to enquire about hosting a science blog on Sciblogs, contact the .
Sciblogs will be live on Wednesday September 30 and you will be able to keep tabs on our Scibloggers via RSS, email updates and by following Sciblogs on Twitter.
Also – notice how many more commenters here have their own avatar. I think that's something to do with their involvement in Sciblogs.co.nz
Related articles by Zemanta
- NZ science bloggers – new opportunity (openparachute.wordpress.com)
- Science communication in New Zealand (openparachute.wordpress.com)
Depressed? Anxious? Aren't we all? Sep 25No Comments
I'm currently reading Zoltan Torey's book The Crucible of Consciousness: An Integrated Theory of Mind and Brain. It's fascinating and I will put up a review some time soon.
In the model he proposes for the self aware mind (consciousness) he deals with problems the mind has to confront. Reflective awareness can lead to chronic anxiety, fretting and anticipation of danger. Human reflective awareness has given us unique and powerful abilities but they "are not altogether a blessing. Or at least blessing that have to be paid for very dearly indeed."
Humans need "neurotic defences" to maintain mental equilibrium. One of these is the fashioning of belief systems – "psychoreligious constructions." These make it difficult for our minds to represent objective reality. Rather they work to maintain our inner peace and sense of security. Our brain is strongly compromised as a data-processing instrument. We also have strong suppression mechanisms which help filter out "ego-threatening stimuli."
The very abilities which have made us an intelligent species actually impair our reason. Rather than a rational species we are a rationalising one. This realisation is extremely important to the scientific endeavour – underling the importance of utilising evidence and testing our ideas against reality, rather than ideology.
The anxieties, obsessions and anticipation of danger can work together with the creation of erroneous belief systems and suppression mechanism to disrupt our mental health. I think we are now starting to realise that mental illness is much more common than we might like to admit. It's more "normal." And this model suggests why.
Futurity recently reported that people vastly underestimate and "underreport the amount of mental illness they've suffered when asked to recall their history years after the fact." This came out of a "long-term tracking study of more than 1,000 New Zealanders from birth to age 32″ (see Depressed? Anxious? Aren't we all?).
'If you start with a group of children and follow them their whole lives, sooner or later almost everybody will experience one of these disorders,' says Terrie Moffitt, the Knut Schmitt-Nielsen professor of psychology and neuroscience at Duke University.
Because self-reporting from memory is the basis for much that is known about the prevalence of mental disorders, anxiety, depression, and substance dependency may actually be twice as high as previously believed, according to new research."
I guess we have come a long way as a society in thinking about mental illness. But we still have a way to go. If we can realise how mental illness arises naturally out of our unique possession of reflective awareness, of consciousness, perhaps we can be nmore acceptable of poeple who suffer from it. After all, none of us is immune. it's very likely we will have bouts of such illness sometime in our lives.
Get in the sack! Sep 22No Comments
Whoops! I know this has a slightly risque meaning in New Zealand. But I am using it here in the way that Irish comedian Dara O'Briain does. It's what he suggests for all those who carp on about the supposed deficiencies of science.
Have a watch of these brief videos to get the full meaning.
Dara O'Briain — If science knew all the answers it would stop!
Dara O'Briain and Frankie Boyle on religion and creationism
Theistic evolution? Sep 21No Comments
This term gets used a lot – but what does it mean?
In a recent discussion a local supporter (I think) of theistic evolution put it this way: Both "theistic evolutionists" and "atheistic evolutionists" accepts Darwinian evolution as true. Nevertheless – he describes these as two alternatives "theories." But he admits: "the empirical evidence . . . will not provide reasons for one position over another. The two positions have to be decided then on other grounds."
I think this person, and probably most other people who use the "theistic evolution," label are confused. They are not talking about scientific theories. They are talking about their own religious beliefs. These "other grounds" are religion.
All these people are saying is: "I accept evolutionary science but I am still a theist."
But why do that? There is only one reality and scientific theories are tested against that reality, not against religious books or opinions of religious leaders. So why attach one's religious beliefs to your acceptance of scientific knowledge?
The fact is that only (some) theists feel the need to declare their religious beliefs in this way. We never hear people saying they accept "atheistic evolution," do we? So it must be something to do with the religious community these theists inhabit.
I think this must arise out of the hostility towards evolutionary science, and sometimes science in general, common in many theistic circles. Certainly, in New Zealand the 20% of the population who reject evolutionary science are mostly Christian – and they comprise about 40% of the local Christian community.
Apparently someone who declares their acceptance of evolutionary science can sometimes be confronted with a very hostile reaction. They are probably considered "sinful" by many fundamentalists. I have heard of cases where such people are shouted down. Even where some families express concern about the children learning about science at school!
So perhaps it's understandable that in this bullying atmosphere many Christians feel the need to add the "theist" adjective to evolutionary. But why give in to such bullying? In the long run bullies are only defeated by standing up to them.
When one gives in and resorts to calling oneself a "theistic evolutionist" isn't it being cowardly?


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Mayor Williams joins 93.7-FM
HUNTINGTON -- Newly-elected Huntington Mayor Steve Williams has agreed to join 93.7-FM The Dawg's morning show with Chuck Black and Virginia West every Tuesday at 8:45 a.m.
West is calling the segment "Talking Smack with Mayor Steve!"
Joining Williams and West during the broadcast is former Huntington mayor and Supertalk 94.1 talk-show host Jean Dean. Dean's words of advice for Mayor Williams are "keep a sense of humor."
Williams laughed at the advice and responded, "I thought she was going to say don't blow it."
The program debuts Tuesday, Jan. 15. The Dawg is owned by Kindred Communications. For more info or to listen live, visit http://kindredcom.net.
The Herald-Dispatch welcomes your comments on this article, but please be civil. Avoid profanity, obscenity, personal attacks, accusations of criminal activity, name-calling or insults to the other posters. Herald-dispatch.com does not control or monitor comments as they are posted, but if you find a comment offensive or uncivil, hover your mouse over the comment and click the X that appears in the upper right of the comment. If you do not want your comment to post to your personal Facebook page, uncheck the box below the comment.


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Hold down Control (PC) or Command (Mac) key + mouseclick to select more than one option
|Avg. Dinner Entrée
|Greater than $25
|$18.01 - $25
|$12.01 - $18
|$7.01 - $12
|Less than $7
Posted by Actual customer on 01/23/2008
The food is ok, with ok service... Mediocre is the only word to describe the place there are better bars in the neighborhood and better places to eat. Though, they do occasionally have good drink specials at early hours, which make it worth eating your dinner there.
Posted by UES on 05/23/2007
I think the below reviews are a bit "fishy" - this place is great and has great specials almost every night but it is not, unfortunately, always hopping. Two burgers or sandwiches for the price of one on Tuesday, and half price apps on Wed are the best. Only get the chicken fingers at half-price, the portion is skimpy. 3 types of fries keeps everyone happy, and the sliders and potato skins are really good. And I don't even like potato skins...
Posted by Fantastic Food, Drink, Trivia & Events on 03/15/2007
This place is always hopping! They have 2-for-1 specials on food and drink, play NTN Trivia, stop in for comedy night, always having something going on. They even have DJ on the weekend and all the college games. The chicken bomb sandwich is to die for and the chicken parmagiana is fantastico too! The weekday bartender is such a doll, he's a really great guy! You should come by and try the cuisine and have a pop with the crew.
Posted by Two for tuesday on 09/17/2006
Excellant place..... fun and young crowd.... cheap drinks and food


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I have been told they are working on the registration site and may be working within 48 hours, but don't quote me on that!
If you are in CA and coming to the SD walk, feel free to send me an email that you are attending the walk and I will be sure to note t-shirt sizes for you. Our t-shirt deadline has been extended to April 11th, which is good news!


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Okay, my OM-4 has been shelved for several years now-in need of a serious CLA, so I've been using my OM-1, today the film advance lever failed to return on its own(sigh), it's been a little sluggish, and feels like it needs a lube job. How hard is it to replace the spring, and perform a home lube? My new(to me)OM-1 hasn't arrived yet, i'm growing a tad impatient to handle it.
Sheesh, I'm getting the blues. I might have to pick my Contax up for a while to satisfy the 35mm needs.
It depends on why the lever didn't return and is it all locked up now? OMs get into an "overwound" condition. Sometimes an experienced user can take off the bottom plate and fiddle with one of several levers and unstick it. But this is usually an indication that the camera needs a CLA, as even if you do unstick it, the problem will probably return. And if you don't have a good idea of just what exactly every gear and lever does, you won't be able to service it yourself. When you remove the bottom plate it's a little intimidating what you'll see in there. It's packed with gears, levers and springs.
My advice is to bite the bullet and just send both camera bodies off to John at Zuiko.com When they return you won't believe how smooth they are and will give you many more years of service.
The lever hasn't been over wound, it just wont return of its own volition, and is somewhat sluggish, like the grease is stiff. Other than that it still functions, its just a PITA to return it manually. I've loked at the exploded view drawing, and it appears the return spring is under the top plate, and beyond my capabilities. I suppose I could just buy a winder and not fuss with it for now.
Well, I put a winder-2 on my OM-1, works just fine, save for one slightly annoying thing, the stupid film advance lever swings out when the motor winds the film. The danged thing smacked me on the eyebrow and startled me. I wrapped a stiff rubber band around it to hold it in position, so far it works.
Be careful, you could put an eye out


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Dutch Crossing, established in 1977, is a peer-reviewed interdisciplinary scholarly journal, devoted to all aspects of Low Countries studies: history and art history of the Low Countries, Dutch and Flemish (and occasionally Afrikaans) literary and cultural studies, linguistics of Dutch and Dutch as a foreign language, and intercultural and transnational studies. Coverage includes both the Netherlands and Belgium, as well as other places where Dutch historically had or continues to have an impact, including parts of the Americas, Southern Africa and South-East Asia. A special focus concerns relations between the Low Countries and the English-speaking world in all periods from the Middle Ages to the present day. Thematic issues have been produced on such topics as Anglo-Dutch relations in the 17th Century; Williamite Scotland and the Dutch Republic; contemporary Dutch women writers; Frisian culture; Landscape Painting; Literary Translation and Medieval Drama. Dutch Crossing aims to encourage research and intellectual exchange, to disseminate scholarly work by younger as well as established researchers, and to enhance the profile of Low Countries Studies and of Dutch and Flemish culture in the English-speaking world. All articles are in English and double-blindly peer-reviewed by at least one referee.
Vol. 39, No. 1 (March 2015)
Table of Contents
|Ulrich Tiedau
|The Royal Households in the Habsburg Netherlands after the Departure of the Household of Burgundy: From the Entourages of the Governors-General to the Maison Royale de Bruxelles
|José Eloy Hortal Muñoz, Koldo Trápaga Monchet
|The Légion Hollandaise d'Orange: Dynastic networks, coalition warfare and the formation of the modern Netherlands, 1813–14
|Mark Edward Hay
|The use of history in Belgium and the Netherlands, 1946–1965: Presentism and Historicism in the Work of Jan Romein, Pieter Geyl and Leopold Flam
|Jo Tollebeek
|Dutch printer and publisher Joan Blaeu II (1596–1673) identified as the subject of a portrait by Michiel van Musscher
|Robert E. Gerhardt
|J. Bernlef and American poetry: Poetic-strategic mentions of Marianne Moore. A case study
|Yves T'Sjoen
|Evan Haefali – New Netherland and the Origins of Ameri-can Religious Liberty, Philadelphia: University of Philadelpha Press, 2012
|Oliver Finnegan
|Raingard Esser: The Politics of Memory: The Writing of Partition in the Seventeenth-Century Low Countries. Brill's Studies in Intellectual History 208. Leiden and Boston: Brill, 2012. ISBN 978-9004208070
|Eddy Verbaan
|Luc Rombouts: Singing Bronze: A History of Carillon Music, Leuven: Lipsius, 2014
|Jeroen Dewulf
Dutch Crossing: Journal of Low Countries Studies – UCL Dept. of Dutch – Gower Street – London WC1E 6BT – UK – +44 (0)20 7679 7756 – © 1977–2014Disclaimer | Freedom of Information | Accessibility | Privacy | Cookies | Advanced Search | Contact Us


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Carley McMaster never gave up – not after a brain tumor and a total of five strokes.
Three years after Carley lost her battle, her parents haven't given up either.
Scott and Vindi McMaster hosted their second "A Dinner of Wine and Roses" benefit and auction Friday – raising funds for their own Carley's Rays of Hope nonprofit in conjunction with the Atlanta-based Brain Tumor Foundation. The organization provides a number of benefit opportunities, including a 5K/10K run coming in June.
The program named after Carley provides assistance through the foundation and much more to help families with children who are being treated for brain tumors.
Special guests of the McMasters at the gala were Chris and Elisha Thompson of Evans, Ga., and their three children. Their son, Christopher Jr., 7, was diagnosed with a tumor last July. Since then, he has undergone three brain surgeries and 24 chemotherapy treatments.
His attitude is unreal, Chris Thompson said. His son remains happy and never gets upset with all the issues he has experienced.
When the family heard about Carley's Rays of Hope, they were connected to the foundation, which has been a huge help with basic needs.
"It means a lot to be here and that Chris can be a part of it," Elisha said.
Vindi expressed her joy that Christopher, his parents and two sisters attended the event, then presented Elisha with flowers.
Carley spent eight months as an out-patient at Hitchcock Healthcare – the rehabilitation facility in Aiken. The McMasters presented its administrators and therapists with the Ray of Hope Award in gratitude – not just for Carley, they said, but for all the thousands of children that Hitchcock has served.
A year ago, pediatrician Dr. John Tiffany received the award for his extraordinary support of children and their health. On Friday, he presented the same award to Heather Raynack, Hitchcock's rehabilitation services director, and recognized the McMasters, too.
"Carley's Rays of Hope has a special place in my heart," Tiffany said. "When the organization was formed three years ago, I saw the courage of Vindi and Scott to give back to the community after going through the loss of a child."
Tiffany called the award last year the highlight of his career. He is also proud of Hitchcock, serving most recently with the agency by meeting with the pediatric team to discuss therapy issues.
"Hitchcock is more than numbers with the therapy. ... It's love and caring," Tiffany said.
Hitchcock staffers constantly meet parents who want their children to eat and get nutrition or learn to walk or say "mommy" or be strong enough to go to school, Raynack said.
"Those basic things are what push us to become excellent therapists," she said. "Our patients become our children, and we want the best for them, too. Our compassion, care and commitment to these children push us to always do more because, quite frankly, they deserve it."
Since the establishment of Carley's Rays of Hope, "We could never have imagined where it's gone," Scott McMaster said. "When we first started, we said we would do one event. Now we're constantly thinking of new things to do to bring in more money for these parents."
STAFF PHOTO BY ROB NOVIT Joined by older sister Brook, Christopher Thompson Jr. was a special guest at the Carley's Rays of Hope benefit gala Friday. Christopher maintains an ongoing smile - despite enduring three brain surgeries and 24 chemotherapy treatments in the last six months.×
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Dartblog Monthly Archives
This is an archive page. Please click here to see the latest entries.
July 2014
Sunday, July 6, 2014
Good to see that Phil Hanlon is out and about in Hanover. We spotted him this week standing in line at Morano Gelato waiting to order ice cream. His wife Gail Gentes is standing behind…
Featured posts
August 14, 2013
Breaking: Of Crips and Bloods and Memories of Ghetto Parties
History repeats itself, first as tragedy and then as farce, or sometimes it just repeats itself. From the New York Times on November 30, 1998: At Dartmouth College, white students at a "ghetto party" dressed…
June 25, 2013
Dean of the College Charlotte Johnson's War on Students Part (2/2)
Part 1, Part 2 Today's post again recounts the events that befell the Freshman. However, the content of the Hanover Police department report reproduced in this space yesterday is supplemented by information from my own…
October 18, 2009
When Love Beckoned in 52nd Street
We were at San Francisco's BIX last evening, enjoying prosecco, cheese, and a bit of music. A full year of inhabitation in Northern California has unraveled to me no decent venue for proper lounging, but…
October 9, 2009
D Afraid of a Little Competish
So our colleague and Dartblog writer Joe Asch informed me that the D has rejected our cunning advertising campaign. Uh-oh. The Dartmouth is widely known as a breeding ground for instant New York Times successes,…
September 4, 2009
How Regents Should Reign
As Dartmouth alumni proceed through the legal hoops necessary to defuse a Board-packing plan—which put in unhappy desuetude an historic 1891 Agreement between alumni and the College guaranteeing a half-democratically-elected Board of Trustees—it strikes one…
August 29, 2009
Election Reform Study Committee
If you are an alum of the College on the Hill, you may have received a number of e-mails of late beseeching your input for a new arm of the College's Alumni Control Apparatus called…


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United have already turned down one bid from Chelsea for the 27-year-old, who is unsettled at Old Trafford.
Chelsea boss Jose Mourinho has stated that Rooney is the Blues only current transfer target and it is expected the Portuguese will lodge another bid for the player.
However, United's stance regarding the England international has not changed.
And they expect the hamstring injury that forced him to leave United's pre-season tour to have healed in time for Rooney to be in Stockholm for the friendly against AIK Solna on 6 August.
"We have said and the club have said he is not for sale," said Moyes after Tuesday's 3-2 friendly defeat against Yokohama F. Marinos. "That has not changed.
"I do expect Wayne to be in Stockholm. As far as I know, he is coming on.
"He has started light jogging and the feedback from the physios is that he is progressing, so hopefully those dates will still be right when we get back."
Pleasing effort
In Rooney's absence, United have struggled during pre-season with J. League side Yokohama F. Marinos inflicting their second loss in three tour games.
Behind after just 27 seconds, United turned the game around through a Jesse Lingard strike and Masakazu Tashiro's own goal.
But the home side hit back to equalise before Yoshihito Fujita bagged the winner three minutes from time.
"I am disappointed with the goals we conceded," said Moyes.
"We had a lot of opportunities to finish the game off at 2-1 and even after that, we had chances which we didn't take.
"But the biggest thing about tonight was the effort the players had to put in.
"We tried to get as many of them through 90 minutes as we could and they worked really hard.
"Some of them did very well and I am pleased with that."
Robin van Persie came off at half-time, while Danny Welbeck picked up a groin strain, but Moyes played down any concerns about the two players.
"Robin just felt his thigh in the first half and we didn't want to take any chances," he said.
"Danny Welbeck felt his groin when he came on and we've just got to make sure they're all okay especially with so many games coming up.
"We've still got a few games to play before the season starts when we get back home."


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Song List for Snap #230 "Fight of Your Life"Submitted by snapjudgment on Fri, 09/20/2013 - 16:14
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Song List for Snap #415 "The Reunion"Submitted by snapjudgment on Fri, 07/12/2013 - 18:10
Song List for Snap #409 "Outside Looking In"Submitted by snapjudgment on Thu, 05/02/2013 - 02:16
Full List of Songs - "Making It Work"Submitted by snapjudgment on Sat, 03/16/2013 - 18:38
Full List of Songs - A Love SupremeSubmitted by stephaniesays on Tue, 02/12/2013 - 14:57
Hey there, you handsome devil. Yeah, we know you want it--the full list of songs from our Valentine's Day ep, "A Love Supreme." xoxo!
Full List of Songs - The CrownSubmitted by stephaniesays on Fri, 01/25/2013 - 15:35
Full List of Songs - SurvivorSubmitted by stephaniesays on Fri, 01/25/2013 - 15:28
I know you're thirsty for some new tunes. Let Snap help you out. Listen to our Survivor episode, then check our your favorite songs right here.
The Large Professor Amaman
Song List for Snap #401 "The Stranger"Submitted by stephaniesays on Fri, 01/25/2013 - 15:20
Strange no more, now the full list of songs for the Stranger will help you find your fav songs!
Full List of Songs - Partner in CrimeSubmitted by stephaniesays on Tue, 11/20/2012 - 15:17
We are your friends. You will never be alone again. So come on, take a look at our Partner in Crime full list of songs!
Intros / Outros
Full List of Songs - The Great OutdoorsSubmitted by stephaniesays on Wed, 11/07/2012 - 14:11
Indeed, good people. We got all your songs from The Great Outdoors episode right here. Roar!
Intros / Outros
Homeboy Sandman - Sputnik
Celph Titled - I could Write A Rhyme: The Instrumentals
9th Wonder & Buckshot - Stop rapping


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Drug Compare
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- Basic Info
Generic: permethrin topical
It skin cream is used to treat scabies.
What if I miss a dose?This does not apply.
What may interact with this medicine?Interactions are not expected. Do not use any other skin products on the affected area without telling your doctor or health care professional.
What should I watch for while using this medicine?It is not unusual for itching and rash to continue for as long as 2 to 4 weeks after treatment. These symptoms may be a temporary reaction to the remains of the mites. This does not mean this cream did not work or that it needs to be reapplied. If you feel that the itching and rash is intense or if it continues beyond 4 weeks, talk to your doctor or health care professional right away.
Scabies is spread by direct skin contact with an infected person. Family members and sexual partners may require treatment with this medicine. You should discuss this with your doctor or health care professional.
Using a normal washing cycle, you should wash all clothing, towels and bed linen that has touched your skin. You do not need to rewash clean clothing that has not yet been worn. Coats, furniture, rugs, floors, and walls do not need to be cleaned in any special manner.
What side effects may I notice from receiving this medicine?Side effects that usually do not require medical attention (report to your doctor or health care professional if they continue or are bothersome):
Where should I keep my medicine?Keep out of the reach of children.
Store at room temperature away from heat and direct light. Do not refrigerate or freeze. Throw away any unused medicine after the expiration date.
Remember, keep this and all other medicines out of the reach of children, never share your medicines with others, and use this medication only for the indication prescribed.
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