Sodium amobarbital: historical perspectives and neurorehabilitation clinical caveats.

The sodium amobarbital (amytal) (SA) interview is a technique that has been utilized in the treatment of a variety of disorders since its introduction in 1929. Since that time, there has been an assortment of research conducted showing its value in both differential diagnosis and treatment of multiple conditions. Notwithstanding the substantive amount of experience with the technique and its application to a myriad number of clinical conditions, it remains a seldom used procedure in clinical practice and certainly in neurorehabilitation. This paper will review the history of SA, as well as summarize the literature published over the past two decades on the clinical applications of SA to provide readers with a foundation for the utility of this agent, as well as the sodium amytal interview (SAI) in neurorehabilitation clinical practice. Special emphasis will be placed on the use of the SAI in individuals with functional disorders that may be seen in the neurorehabilitation setting, as well as various classes of pain disorders.

[The use of enzymatic hydrolysis for isolation of barbituric acid derivatives from blood (as exemplified by phenobarbital and barbamyl)].

Modern isolation techniques by direct extraction with organic solvents or after protein precipitation by various sedimenting or salting-out agents are characterized by low efficiency and do not permit to liberate derivatives of barbituric acid from their complexes with blood proteins. The use of enzymatic hydrolysis makes it possible to break bonds between barbiturates and protein and thereby improve the efficiency of isolation. We performed enzymatic hydrolysis of the model phenobarbital-blood and barbamyl-blood complexes with the use of trypsin, pepsin, chymotrypsin, and papain. The degree of phenobarbital extraction with trypsin and barbamyl was estimated at 62.1 +/- 1.2% and 75.1 +/- 1.6% respectively; in other words, it was 32.7 +/- 1.0% and 51.1 +/- 1.0% higher than that achieved by traditional methods. Certain validation characteristics of the new method are presented.

Molecular mechanisms and binding site location for the noncompetitive antagonist crystal violet on nicotinic acetylcholine receptors.

We investigated the molecular mechanisms and the binding site location for the fluorophor crystal violet (CrV), a noncompetitive antagonist of the nicotinic acetylcholine receptor (AChR). To this end, radiolabeled competition binding, fluorescence spectroscopy, Schild-type analysis, patch-clamp recordings, and molecular dynamics approaches were used. The results indicate that (i) CrV interacts with the desensitized Torpedo AChR with higher affinity than with the resting state at several temperatures (5-37 degrees C); (ii) CrV-induced inhibition of the phencyclidine (PCP) analogue [(3)H]thienylcyclohexylpiperidine binding to the desensitized or resting AChR is mediated by a steric mechanism; (iii) tetracaine inhibits CrV binding to the resting AChR, probably by a steric mechanism; (iv) barbiturates modulate CrV binding to the resting AChR by an allosteric mechanism; (v) CrV itself induces AChR desensitization; (vi) CrV decreases the peak of macroscopic currents by acting on the resting AChR but without affecting the desensitization rate from the open state; and (vii) two tertiary amino groups from CrV may bind to the alpha1-Glu(262) residues (located at position 20') in the resting state. We conclude that the CrV binding site overlaps the PCP locus in the resting and desensitized state. The noncompetitive action of CrV may be explained by an allosteric mechanism in which the binding of CrV to the extracellular mouth of the resting receptor leads to an inhibition of channel opening. Binding of CrV probably increases desensitization of the resting channel and stabilizes the desensitized state.

Interaction of barbiturate analogs with the Torpedo californica nicotinic acetylcholine receptor ion channel.

Barbiturate-induced anesthesia is a complex mechanism that probably involves several ligand-gated ion channel superfamilies. One of these superfamilies includes the archetypical nicotinic acetylcholine receptor (nAChR), in which barbiturates act as noncompetitive antagonists. In this regard, we used the Torpedo californica nAChR and a series of barbiturate analogs to characterize the barbiturate binding site(s) on this superfamily member. [(14)C]Amobarbital binds to one high-affinity (K(d) = 3.7 microM) and several (approximately 11) low-affinity (K(d) = 930 microM) sites on the resting and desensitized nAChRs, respectively. Characteristics of the barbiturate binding site on the resting nAChR include: (1) a tight structure-activity relationship. For example, the barbiturate isobarbital [5-ethyl-5'-(2-methylbutyl) barbituric acid] is >10-fold less potent than its formula isomer amobarbital [5-ethyl-5'-(3-methylbutyl) barbituric acid] in inhibiting [(14)C]amobarbital binding. (2) A binding locus within the pore of the nAChR ion channel. Each of the barbiturate analogs inhibited the binding of [(3)H]tetracaine or photoincorporation of 3-trifluoromethyl-3-(m-[(125)I]iodophenyl) diazirine in a mutually exclusive manner. (3) Stereoselective binding. The R(+)-enantiomers of isobarbital and pentobarbital are approximately 2-fold more potent in inhibiting 3-trifluoromethyl-3-(m-[(125)I]iodophenyl) diazirine photoincorporation than the S(-)-enantiomers. Finally, molecular modeling suggests that within the channel, the pyrimidine ring of the barbiturate is located just above the highly conserved leucine ring (M2--9; e.g., delta Leu-265), whereas the 5' side chain projects downward, and depending upon its conformation, introduces steric hindrance to binding because of the restriction in the lumen of the channel introduced by the leucine side chains.

In vitro and in vivo evaluation of an amylobarbitone/hydroxypropyl-beta-cyclodextrin complex prepared by a freeze-drying method.

The complex formation of amylobarbitone (AMB) with 2-hydroxypropyl-beta-cyclodextrin (HP-beta-CD) was investigated in aqueous solution and in the solid state. The apparent stability constant for complex formation (Kc) calculated by phase solubility and spectral shift methods was 524 M-1 and 568 M-1, respectively. The stoichiometric molar ratio of the complex was estimated to be 1:1 and the solubility of AMB in water was increased about 3 fold. The solid dispersion system of AMB/HP-beta-CD in 1:1 molar ratio was prepared by a freeze-drying method. Differential scanning calorimetry (DSC), x-ray diffractometry, (IR) and 1H NMR spectroscopy were used to confirm that inclusion between the drug and HP-beta-CD occurred. The dissolution behavior of the drug as a physical mixture as well as the prepared complex, showed enhanced drug dissolution properties of the prepared complex compared to the physical mixture or the drug alone. The dissolution rate appeared in the first 2 min, 25 times greater for the complex than for the drug alone. Furthermore, in-vivo study revealed that the duration and hypnotic activity of AMB after its oral administration to mice were improved by inclusion.

Anatomic correlates of memory from intracarotid amobarbital injections with technetium Tc 99m hexamethylpropyleneamine oxime SPECT.

OBJECTIVE: To better identify regions of the brain affected by intracarotid amobarbital injections and to more precisely predict whether resections of specific brain regions will cause postoperative memory deficits. DESIGN: We modified the standard intracarotid amobarbital procedure by adding a radioactive tracer to the amobarbital injection, thereby providing better correlation between behavior and deactivated brain region. SETTING: Tertiary-care hospital center with a dedicated program for medical and surgical treatment of epilepsy. PATIENTS: We studied 39 patients with medically intractable epilepsy drawn from a regional referral base. INTERVENTION: Intracarotid injection of 125 mg of sodium amobarbital with 37 MBq of technetium Tc 99m hexamethylpropyleneamine oxime (HMPAO), followed by language and memory testing. MAIN OUTCOME MEASURES: The distribution of amobarbital as measured by single photon emission computed tomographic imaging of HMPAO and patient performance on memory tasks. RESULTS: Medial temporal regions were irrigated by the amobarbital in only 28% of the injections. Overall, findings suggest that medial temporal and lateral neotemporal cortex play a role in memory. CONCLUSIONS: The regions involved in memory function vary by individual, as does the distribution of amobarbital. Thus, the most accurate method of determining correlation of brain region with memory function during intracarotid amobarbital injection involves the use of a tracer such as HMPAO.

[Crystal morphological and habit investigations of crystalline drugs. II. Amobarbital, barbital and ephedrine hydrochloride].

Crystal morphology, habit and refractive indices necessary for the analyses of amobarbital, barbital, phase III and ephedrine hydrochloride were studied using a polarizing microscope and partly using a reflexing goniometer. These substances, market products of Japanese Pharmacopoeia XII, were measured for optical properties including key refractive indices to confirm their modifications, and their possible facial angles along the zone of which axis is parallel to the elongated direction was also measured using a goniometer. Stereographic projections were drawn and the crystal habits of market products were shown by orthographic projections. It was also found that one of the key refractive indices of each modification was specially usable for the analyses of crystal habits including their crushed powders.

Identification of 5-ethyl-5-(2-methylbutyl)barbituric acid as an impurity of manufacture in amobarbital.

Amobarbital [5-ethyl-5-(3-methylbutyl)barbituric acid], USP, was found to contain an impurity that was not associated with hydrolysis and decomposition of the barbiturate ring. The impurity was isolated by semipreparative HPLC and was identified as 5-ethyl-5-(2-methylbutyl)barbituric acid (1) by MS (electron impact and chemical ionization) and 1H NMR. The substitution pattern on the alkyl side chain was verified by using the achiral NMR shift reagent tris(6,6,7,7,8,8,8-heptafluoro-2,2- dimethyl-3,5-octanedionato)europium(III). Older samples of amobarbital, USP, contained greater than 6% of 1, whereas recent samples of amobarbital, USP, contained less than 1% of 1. Because the pharmacological profiles of 1 and amobarbital in rodents are comparable, the impurity probably does not constitute a clinically significant problem for humans.