Compositions and methods for derepressing RE1 silencing transcription factor target genes
Inventors
Nesti, Edmund • Pisarchik, Alexander
Assignees
Alcamena Stem Cell Therapeutics LLC
Publication Number
US-12180255-B2
Publication Date
2024-12-31
Expiration Date
2040-11-20
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Abstract
The invention relates to compounds, compositions, and methods for derepressing RE1 silencing transcription factor (REST) target genes are provided. In particular, a peptide having the sequence TEDLEPPEPPLPKEN (SEQ. ID NO: 1) and EDLEPPEPPLPK (SEQ. ID NO: 15), or the reversed sequences made of D-amino acids (retro inverted, RI) nekplppeppeldet (SEQ ID NO: 16) and kplppeppelde (SEQ ID NO: 17), are disclosed for inhibiting REST activity. The peptides are useful to treat, prevent, or ameliorate conditions such as traumatic brain injury, epilepsy, dementia, Huntington's Disease (HD), chronic pain, brain cancer (including glioblastoma multiforme), pancreatic cancer; diabetes, and peripheral nerve injury.
Core Innovation
The invention provides compounds, compositions, and methods for derepressing RE1 silencing transcription factor (REST) target genes by inhibiting REST activity. The core innovation includes peptides having the sequences TEDLEPPEPPLPKEN (SEQ ID NO: 1), EDLEPPEPPLPK (SEQ ID NO: 15), or their reversed sequences made of D-amino acids (retro inverted, RI) nekplppeppeldet (SEQ ID NO: 16) and kplppeppelde (SEQ ID NO: 17), which bind to C-terminal domain small phosphatase 1 (CTDSP1) to inhibit REST activity.
The problem solved by the invention arises from the role of REST as a repressor of hundreds of neuronal genes essential for terminal neuronal differentiation and function, including ion channels and growth factors. Persistence of REST impedes terminal neuronal differentiation, neuronal survival, and nerve regeneration. REST is stabilized by CTDSP1 through dephosphorylation, preventing its degradation and thus maintaining gene repression. Therefore, a method to inhibit CTDSP1 and promote REST degradation is needed to allow expression of REST target genes and facilitate neuronal differentiation and regeneration.
The invention discloses REST phosphomimetic peptides (RPPs) and 98 REST phosphomimetic peptide variants (RPPVs) that bind CTDSP1 with high affinity to inhibit its phosphatase activity on REST, leading to REST degradation. Fusion of RPPs or RPPVs to intracellular transport peptides, such as cell penetrating peptides (CPPs) and endosomal release sequences, and their cyclization further improve intracellular transport, binding affinity, and stability. These peptides promote expression of REST target genes, neural differentiation, and are useful to treat diseases or conditions associated with REST and CTDSP1.
Claims Coverage
The patent contains three main independent claims relating to isolated peptides comprising specific amino acid sequences and their pharmaceutical compositions.
Isolated peptides comprising specified amino acid sequences
Isolated peptides comprising amino acid sequences selected from SEQ ID NOS: 1, 15-17, 18, 22, 25, 31, 35, 60, 63, 64, 66, 68, 75, 81, 85, 110, 113, 114, and 116.
Peptides fused to cell penetrating peptides or endosomal release sequences
Peptides as defined above that are fusion peptides fused to cell penetrating peptides or endosomal release sequences, specifically those selected from SEQ ID NOS: 118-137 or 140-159, optionally connected via linkers from SEQ ID NOS: 138, 139, 160, and 161, fused at N- or C-terminus.
Cyclized fusion peptides with improved properties
Isolated peptides comprising cyclized fusion peptides selected from SEQ ID NOS: 2, 5, 12, 13 and 14 that are fused to cell penetrating peptides or endosomal release sequences at N- or C-terminus.
The claims cover isolated REST phosphomimetic peptides and variants that bind CTDSP1 to inhibit REST activity, including fusion peptides with intracellular transport sequences and their cyclized forms, as well as pharmaceutical compositions containing these peptides for therapeutic use.
Stated Advantages
High binding affinity of the REST phosphomimetic peptides to CTDSP1 (low picomolar) with slow off rate suggests low risk of off-target effects.
The peptides promote REST degradation, leading to expression of neuronal genes, neuronal differentiation, and nerve regeneration.
Cyclization and use of D-amino acids increase peptide stability and half-life, facilitating therapeutic applicability.
Peptides are cell permeable and localize to the nucleus where REST and CTDSP1 act.
Therapeutic potential for treating diseases involving REST such as traumatic brain injury, chronic pain, epilepsy, neurodegenerative diseases, brain and pancreatic cancers, and diabetes.
Preliminary data indicate that the peptides increase expression of genes suppressed by REST, promote neurogenesis, are not neurotoxic, and can improve functional recovery in relevant models.
Documented Applications
Treatment, prevention, or amelioration of traumatic brain injury.
Treatment, prevention, or amelioration of epilepsy.
Treatment, prevention, or amelioration of dementia and age-related neurodegenerative diseases including Alzheimer's and Huntington's Disease.
Treatment, prevention, or amelioration of chronic pain and peripheral nerve injury.
Treatment, prevention, or amelioration of brain cancers including glioblastoma multiforme.
Treatment, prevention, or amelioration of pancreatic cancer.
Treatment, prevention, or amelioration of diabetes.
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