gpi-5693 and Pain

E2072 [(3-2-mercaptoethyl)biphenyl-2,3'-dicarboxylic acid] is a novel, potent and selective thiol-based glutamate carboxypeptidase II (GCP-II) inhibitor that has shown robust analgesic and neuroprotective efficacy in preclinical models of neuropathic pain and chemotherapy-induced peripheral neuropathy. For the first time, we describe the drug metabolism and pharmacokinetic profile of E2072 in rodents and primates. Intravenously administered E2072 was found to exhibit an unexpectedly long terminal half-life (105 ± 40 h) in rats. The long half-life was found to be the result of its ability to rapidly form reversible homo- and possibly heterodisulfides that served as a continuous E2072 depot. The half-life of reversible homodisulfides was 208 ± 81 h. In further support, direct intravenous administration of the E2072-homodisulfide in rats resulted in the formation of E2072, with both E2072 and its disulfide detected in plasma up to 7 days after dose. The observed long exposures were consistent with the sustained efficacy of E2072 in rodent pain models for several days after dose cessation. It is noteworthy that a shorter t(½) of E2072 (23.0 ± 1.2 h) and its homodisulfide (21.0 ± 0.95 h) was observed in primates, indicating interspecies differences in its disposition. In addition, E2072 was found to be orally available with an absolute bioavailability of ∼30% in rats and ∼39% in monkeys. A tissue distribution study of E2072 and its homodisulfide in rats showed good tissue penetration, particularly in sciatic nerve, the presumed site of action for treatment of neuropathy. Metabolic stability and the correlation between pharmacokinetic profile and pharmacological efficacy support the use of this GCP-II inhibitor in the clinic.

Topics: Animals; Benzoates; Biological Availability; Disulfides; Dogs; Glutamate Carboxypeptidase II; Half-Life; Humans; Macaca fascicularis; Male; Mice; Microsomes, Liver; Pain; Rats; Rats, Sprague-Dawley; Sulfhydryl Compounds; Tissue Distribution

A series of thiol-based inhibitors containing a benzyl moiety at the P1' position have been synthesized and tested for their abilities to inhibit glutamate carboxypeptidase II (GCP II). 3-(2-Carboxy-5-mercaptopentyl)benzoic acid 6c was found to be the most potent inhibitor with an IC(50) value of 15 nM, 6-fold more potent than 2-(3-mercaptopropyl)pentanedioic acid (2-MPPA), a previously discovered, orally active GCP II inhibitor. Subsequent SAR studies have revealed that the phenoxy and phenylsulfanyl analogues of 6c, 3-(1-carboxy-4-mercaptobutoxy)benzoic acid 26a and 3-[(1-carboxy-4-mercaptobutyl)thio]benzoic acid 26b, also possess potent inhibitory activities toward GCP II. In the rat chronic constriction injury (CCI) model of neuropathic pain, compounds 6c and 26a significantly reduced hyperalgesia following oral administration (1.0 mg/kg/day).

Topics: Analgesics; Animals; Antigens, Surface; Benzoates; Chronic Disease; Constriction, Pathologic; Glutamate Carboxypeptidase II; Glutarates; Humans; Pain; Peripheral Nervous System Diseases; Rats; Structure-Activity Relationship; Sulfhydryl Compounds

Excitotoxic glutamate release occurs in several neurological disorders. One source is derived from the hydrolysis of the neuropeptide N-acetyl aspartyl glutamate (NAAG) by glutamate carboxypeptidase II (GCPII, also known as NAALADase). Drugs that attenuate glutamate transmission have been shown to relieve neuropathic pain, however side effects have limited their clinical use. It appears that GCPII is exclusively recruited to provide a glutamate source in hyperglutamatergic, excitotoxic conditions and therefore would be devoid of such side effects. Here we report on the therapeutic effects of an orally bio-available GCP II inhibitor on established painful and sensory neuropathy in the spontaneously diabetic BB/Wor rat. It significantly improved hyperalgesia, nerve conduction velocity and underlying myelinated fiber atrophy. The data suggest that GCP II inhibition may provide a meaningful and effective approach to the treatment of painful diabetic neuropathy.

Topics: Analysis of Variance; Animals; Diabetic Neuropathies; Disease Models, Animal; Female; Glutamate Carboxypeptidase II; Glutarates; Male; Nerve Fibers, Myelinated; Neural Conduction; Pain; Pain Measurement; Rats; Rats, Inbred BB; Reaction Time; Sodium-Potassium-Exchanging ATPase; Sulfhydryl Compounds

Two representative glutamate carboxypeptidase II (GCP II) inhibitors, 2-(hydroxypentafluorophenylmethyl-phosphinoylmethyl)pentanedioic acid 2 and 2-(3-mercaptopropyl)pentanedioic acid 3, were synthesized in high optical purities (>97%ee). The two enantiomers of 2 were prepared from previously reported chiral intermediates, (R)- and (S)-2-(hydroxyphosphinoylmethyl)pentanedioic acid benzyl esters 8. The synthesis of (R)- and (S)-3 involves the hydrolysis of (R)- and (S)-3-(2-oxo-tetrahydro-thiopyran-3-yl)propionic acids, (R)- and (S)-11, the corresponding optically pure thiolactones delivered by chiral chromatographic separation of the racemic 11. GCP II inhibitory assay revealed that (S)-2 is 40-fold more potent than (R)-2. In contrast, both enantiomers of 3 inhibited GCP II with nearly equal potency. The efficacy observed in subsequent animal studies with these enantiomers correlated well with the inhibitory potency in a GCP II assay.

Topics: Analgesics; Animals; Brain Ischemia; Cerebral Cortex; Constriction, Pathologic; Crystallography, X-Ray; Glutamate Carboxypeptidase II; Glutarates; Infarction, Middle Cerebral Artery; L-Lactate Dehydrogenase; Molecular Structure; Neuroprotective Agents; Pain; Peripheral Nervous System Diseases; Phosphinic Acids; Rats; Stereoisomerism; Structure-Activity Relationship; Sulfhydryl Compounds; Tissue Culture Techniques

A series of 2-(thioalkyl)pentanedioic acids were synthesized and evaluated as inhibitors of glutamate carboxypeptidase II (GCP II, EC 3.4.17.21). The inhibitory potency of these thiol-based compounds against GCP II was found to be dependent on the number of methylene units between the thiol group and pentanedioic acid. A comparison of the SAR of the thiol-based inhibitors to that of the phosphonate-based inhibitors provides insight into the role of each of the two zinc-binding groups in GCP II inhibition. The most potent thiol-based inhibitor, 2-(3-mercaptopropyl)pentanedioic acid (IC(50) = 90 nM), was found to be orally bioavailable in rats and exhibited efficacy in an animal model of neuropathic pain following oral administration.

Topics: Administration, Oral; Analgesics; Animals; Biological Availability; Carboxypeptidases; Constriction, Pathologic; Enzyme Inhibitors; Glutamate Carboxypeptidase II; Glutarates; Hot Temperature; Hyperalgesia; Male; Pain; Peripheral Nervous System Diseases; Rats; Rats, Sprague-Dawley; Sciatic Nerve; Structure-Activity Relationship; Sulfhydryl Compounds