RESUMEN
The synthetic derivative of pregnenolone MAP4343 (3ß-methoxy-pregnenolone) binds in vitro to microtubule-associated-protein 2 (MAP2), stimulates the polymerization of tubulin, enhances the extension of neurites and protects neurons against neurotoxic agents. Its efficacy was assessed in vivo with the most commonly used thoracic spinal cord compression/contusion models in rats. In the three models used, the post-traumatic subcutaneous injection of MAP4343 significantly improved the recovery of locomotor function after spinal cord injury, as shown by an earlier and more complete recovery compared to vehicle-treated rats. The first injection of MAP4343 could be delayed up to 24h after spinal cord injury with maintained efficiency. The improvement was correlated with the preservation of both dendritic trees of motoneurons in the lumbar spinal cord caudally to the injury site, and of MAP2 at lesion site and in the lumbar spinal cord. The results obtained in three different rat models of spinal cord injury demonstrate the beneficial effects of this therapeutic strategy and identify MAP4343 as a potential treatment for acute spinal cord injury.
Asunto(s)
Fármacos Neuroprotectores/farmacología , Pregnenolona/análogos & derivados , Traumatismos de la Médula Espinal/tratamiento farmacológico , Animales , Western Blotting , Modelos Animales de Enfermedad , Inmunohistoquímica , Masculino , Microtúbulos/efectos de los fármacos , Pregnenolona/farmacología , Ratas , Ratas Sprague-Dawley , Recuperación de la Función/efectos de los fármacos , Traumatismos de la Médula Espinal/patologíaRESUMEN
The neurosteroid pregnenolone (PREG) and its chemically synthesized analog 3beta-methoxypregnenolone (MePREG) bind to microtubule-associated protein 2 (MAP2) and stimulate the polymerization of microtubules. PREG, MePREG, and progesterone (PROG; the physiological immediate metabolite of PREG) significantly enhance neurite outgrowth of nerve growth factor-pretreated PC12 cells. However, PROG, although it binds to MAP2, does not increase the immunostaining of MAP2, contrary to PREG and MePREG. Nocodazole, a microtubule-disrupting agent, induces a major retraction of neurites in control cultures, but pretreatment with PREG/MePREG is protective. Decreasing MAP2 expression by RNA interference does not modify PROG action, but it prevents the stimulatory effects of PREG and MePREG on neurite extension, showing that MAP2 is their specific receptor.
Asunto(s)
Proteínas Asociadas a Microtúbulos/metabolismo , Microtúbulos/efectos de los fármacos , Neuritas/efectos de los fármacos , Pregnenolona/análogos & derivados , Pregnenolona/farmacología , Receptores de Esteroides/metabolismo , Animales , Cinética , Masculino , Proteínas Asociadas a Microtúbulos/agonistas , Proteínas Asociadas a Microtúbulos/genética , Microtúbulos/metabolismo , Factor de Crecimiento Nervioso/farmacología , Neuronas/efectos de los fármacos , Neuronas/metabolismo , Neuronas/fisiología , Nocodazol/farmacología , Células PC12 , Progesterona/farmacología , Interferencia de ARN , Ratas , Ratas Sprague-Dawley , Receptores de Esteroides/agonistasRESUMEN
MAPREG (microtubule-associated protein/neurosteroidal pregnenolone) is a start-up company that was created in October 2000. Its acronym recalls the basic discovery (Murakami et al., 2000) from which drug(s) will hopefully be developed that are useful for neuroprotection and repair in conditions such as post-traumatic and postischemic lesions, as well as defects proper to normal aging and neurodegenerative diseases, that is, principally Alzheimer's disease. Pregnenolone, the main steroid synthesized from cholesterol in the nervous system (therefore, a neurosteroid), binds specifically with high affinity (> or = 40 nM) to microtubule-associated protein 2 (MAP2), a protein family involved in the assembly and stabilization of microtubules made from tubulin alpha and beta polymers, and in the bundling of several microtubules by MAP2 projection arms. Pregnenolone binding increases MAP2-induced microtubule polymerization, when purified tubulin and MAP2 are coincubated in GTP containing buffer at 37 degrees C. Therefore, MAP2 can be considered as a receptor for a novel mechanism of steroid action. The underlying principle and its potential pharmacological consequences are described in an INSERM patent (FR 0003430; March 17, 2000). MAPREG has established its own laboratory in a space rented to Bicêtre hospital, near the research building of INSERM, where two of the main founders of the company (Drs. E. Baulieu and P. Robel) work. The company has been quite successful, largely thanks to the support of ISOA (attributed in October 2002). A lead compound (pregnenolone derivative) was tested and patented by MAPREG early in 2003 (FR 0300507; January 17, 2003). Activities and results reported at the ISOA meeting on Oct. 2, 2003, include in vitro basic studies, in vitro and in vivo neuroprotection trials in rodent systems, and studies with human cells and an AD transgenic mouse model.