Protein induced by deprenyl

Details Protein induced by deprenyl Protein induced by deprenyl

1998-08-13

2001-05-15

Spector, Lorraine (Department: 1645)

Chemistry: molecular biology and microbiology

Animal cell, per se ; composition thereof; process of...

C435S320100, C435S252300, C536S023500, C530S350000

Reexamination Certificate

active

06232118

ABSTRACT:

The present invention relates to a novel polypeptide which is induced by a neuroactive drug, to sequences encoding this polypeptide and to uses thereof.
Damaged cells, cells receiving inadequate trophic support or conflicting signals, or deprived of their targets eliminate themselves by means of programmed cell death or apoptosis, leading to their clean demise, leaving no traces. Evidence is accumulating that neuronal cell death in many neurodegenerative diseases including Alzheimer's, Parkinson's, Huntington's disease, amyotrophic lateral sclerosis, cerebellar degeneration, and oligodendrocyte death in multiple sclerosis, is associated with apoptosis. Rescuing cells from apoptosis may halt progression of neurodegenerative diseases, provided the rescued cells remain functional.
(−)—Deprenyl® (Formula 1; referred to herein as Deprenyl®) delays the progression of Parkinson's disease and rescues neurones from presumably apoptotic cell death in a number of in vivo paradigms (death of facial motoneurones after axotomy; destruction of dopaminergic cells in substantia nigra by MPTP or MPP+; death of CA
1
hippocampal neurones after unilateral carotid occlusion followed by a brief period of hypoxia; death of hippocampal pyramidal cells after systemic kainate administration; death of retinal ganglion cells after optic nerve crush) as well as in vitro in trophic factor deprived PC12 cells and oligodendrocytes by an as yet unidentified mechanism.
Deprenyl® is metabolised via (−)—desmethyideprenyl to (−)—methamphetamine and (−)—amphetamine, both of which potently antagonise Deprenyl®'s rescuing effects in vivo and in vitro. Therefore, compounds with equal or better neurorescuing properties than Deprenyl®, but unable to generate antagonistic metabolites, may show a higher therapeutic efficacy in the treatment of neurodegenerative diseases.
We have now found that Deprenyl® induces a novel polypeptide in neural cells. This polypeptide has the structure given in SEQ ID No. 1 and is referred to herein as DIP 1 (Deprenyl® Induced Protein 1).
Accordingly, we provide DIP 1, which preferably has the structure shown in SEQ ID No. 1. The recited sequence is that of rat DIP 1, but the invention relates to DIP 1 derived from all sources, including human DIP 1. Species homologues of rat DIP 1 may be obtained using the recited sequence according to standard methodology, as set out below.
The invention also includes functional derivatives of the polypeptide of SEQ ID No. 1.
“Functional derivative” means that the derivative in question at least one functional determinant of DIP 1. Such functions include the susceptibility to induction by Deprenyl®, and/or at least one in vivo function of DIP 1. For example, derivatives of DIP 1 according to the invention may be able to inhibit or postpone apoptotic neural degeneration. Thus, DIP 1 as provided by the present invention includes splice variants encoded by mRNA generated by alternative splicing of a primary transcript, amino acid mutants, posttranslational modifications, such as glycosylation and phosphorylation variants, and other covalent derivatives of DIP 1 which retain the physiological and/or physical properties of DIP 1. Exemplary derivatives include molecules wherein the protein of the invention is covalently modified by substitution, chemical, enzymatic, or other appropriate means with a moiety other than a naturally occurring amino acid. Such a moiety may be a detectable moiety such as an enzyme or a radioisotope, or other detectable labels, tags, toxins and genes such as oncogenes and tumour suppressor genes. Further included are naturally occurring variants of DIP 1 found within a particular species, preferably a mammal. Such a variant may be encoded by a related gene of the same gene family, by an allelic variant of a particular gene, or represent an alternative splicing variant of the DIP 1 gene.
Derivatives which retain common functional determinants can be fragments of DIP 1. Fragments of DIP 1 comprise individual domains thereof, as well as smaller polypeptides derived from the domains. Preferably, smaller polypeptides derived from DIP 1 according to the invention define a single functional activity which is characteristic of DIP 1. Fragments may in theory be almost any size, as long as they retain one characteristic of DIP 1. Preferably, fragments will be between 5 and 200 amino acids in length. Longer fragments are regarded as truncations of the full-length DIP 1 and generally encompassed by the term “DIP 1”.
Derivatives of DIP 1 also comprise mutants thereof, which may contain amino acid deletions, additions or substitutions, subject to the requirement to maintain at least one feature characteristic of DIP 1. Thus, conservative amino acid substitutions may be made substantially without altering the nature of DIP 1, as may truncations from the 5′ or 3′ ends. Deletions and substitutions may moreover be made to the fragments of DIP 1 comprised by the invention. DIP 1 mutants may be produced from a DNA encoding DIP 1 which has been subjected to in vitro mutagenesis resulting e.g. in an addition, exchange and/or deletion of one or more amino acids. For example, substitutional, deletional or insertional variants of DIP 1 can be prepared by recombinant methods and screened for functional similarity to the native forms of DIP 1.
The fragments, mutants and other derivatives of DIP 1 preferably retain substantial homology with DIP 1. As used herein, “homology” means that the two entities share sufficient characteristics for the skilled person to determine that they are similar in origin and function. Preferably, homology is used to refer to sequence identity. Thus, the derivatives of DIP 1 preferably retain substantial sequence identity with the sequence of SEQ ID No. 1.
“Substantial homology”, where homology indicates sequence identity, means more than 50% sequence identity, preferably more than 75% sequence identity and most preferably a sequence identity of 90% or more.
Preferably, the protein or derivative thereof of the invention is provided in isolated form. “Isolated” means that the protein or derivative has been identified and is free of one OF more components of its natural environment. Isolated DIP 1 includes DIP 1 in a recombinant cell culture. DIP 1 present in an organism expressing a recombinant DIP 1 gene, whether the DIP 1 protein is “isolated” or otherwise, is included within the scope of the present invention.
DIP 1 is believed to be a member of the bcl family of proteins, and is closely homologous to bcl xL. There are, however, characteristic differences between the two proteins which will be apparent by comparing the sequence given herein with the published sequence of rat bcl xL.
The polypeptide according to the invention is closely associated with neurodegenerative disorders and particularly apoptosis in neural cells. Accordingly, the invention provides a composition comprising a polypeptide according to the invention or a modulator thereof for use as a medicament in the treatment or diagnosis of neural diseases.
According to a further aspect of the present invention, there is provided a nucleic acid encoding DIP 1. In addition to being useful for the production of recombinant DIP 1 protein, these nucleic acids are also useful as probes, thus readily enabling those skilled in the art to identify and/or isolate nucleic acid encoding DIP 1. The nucleic acid may be uniabelled or labelled with a detectable moiety. Furthermore, nucleic acid according to the invention is useful e.g. in a method determining the presence of DIP 1-specific nucleic acid, said method comprising hybridising the DNA (or RNA) encoding (or complementary to) DIP 1 to test sample nucleic acid and determining the presence of DIP 1. In another aspect, the invention provides nucleic acid sequence that is complementary to, or hybridises under stringent conditions to, a nucleic acid sequence encoding DIP 1.
The invention also provides a method for amplifying a nucleic acid test sample comprising prim

Affiliated with

Also associated with

Rating

Protein induced by deprenyl does not yet have a rating. At this time, there are no reviews or comments for this patent.

If you have personal experience with Protein induced by deprenyl, we encourage you to share that experience with our LandOfFree.com community. Your opinion is very important and Protein induced by deprenyl will most certainly appreciate the feedback.

Rate now

Comments { 0 }

Profile ID: LFUS-PAI-O-2558046