JPWO2021214443A5 - - Google Patents

Claims (18)

A synthetic CNS-specific promoter or a functional variant thereof comprising a sequence according to any one of SEQ ID NOs: 1-8, 21-26, optionally comprising a sequence that is at least 60%, 65%, 70%, 75%, 80%, 90%, 95%, 96%, 97%, 98% or 99% identical to any one of SEQ ID NOs: 1-8, 21-26, and/or wherein the functional variant of the synthetic CNS-specific promoter retains at least 25%, 50%, 75%, 80%, 85%, 90%, 95% or 100% of the activity of the reference promoter . A CNS-specific cis-regulatory element (CRE) or a functional variant thereof comprising a sequence according to any one of SEQ ID NOs: 9-11, 28-31, optionally comprising a sequence that is at least 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% identical to any one of SEQ ID NOs: 9-11, 28-31. A synthetic CNS-specific cis-regulatory module (CRM) comprising the CRE of claim 2 . A synthetic CNS-specific promoter comprising a CRE according to claim 2 or a CRM according to claim 3 . An isolated minimal or proximal promoter or a functional variant thereof comprising a sequence according to any one of SEQ ID NOs: 12-13, optionally comprising a sequence that is at least 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% identical to any one of SEQ ID NOs: 12-13. 6. A synthetic CNS-specific promoter comprising the minimal or proximal promoter of claim 5 . comprising or consisting of SEQ ID NO:1 or SEQ ID NO:21 or a functional variant thereof , which is broadly active in the brain when administered by intracerebroventricular (ICV) injection, optionally active at a level of at least 100%, 150% or 200% of the activity of synapsin-1 (SEQ ID NO:14) in the brain; or
comprising or consisting of SEQ ID NO:8 or SEQ ID NO:26, which is active in the midbrain and, optionally, active in dopaminergic neurons when administered by intravenous (IV) injection;
2. The synthetic CNS-specific promoter of claim 1 . 10. An expression cassette comprising the synthetic CNS-specific promoter of claim 1, 4, 6 or 7 operably linked to a nucleic acid sequence encoding an expression product. A vector comprising the synthetic CNS-specific promoter of claim 1, 4, 6 or 7 or the expression cassette of claim 8 . 10. The vector of claim 9 , which is a viral vector, such as an AAV vector, an adenoviral vector, a retroviral vector or a lentiviral vector , optionally wherein the vector is a lentiviral vector or an AAV vector. A virion comprising the vector according to claim 10. 12. A pharmaceutical composition comprising the synthetic CNS-specific promoter of claim 1, 4, 6 or 7 , the expression cassette of claim 8 , the vector of claim 9 or 10 or the virion of claim 11 . 13. A synthetic CNS-specific promoter according to claim 1, 4, 6 or 7 , an expression cassette according to claim 8 , a vector according to claim 9 or 10 , a virion according to claim 11 or a pharmaceutical composition according to claim 12 for use as a medicament or for use in the manufacture of a pharmaceutical composition for the treatment of a medical condition or disease. 12. A cell comprising the synthetic CNS-specific promoter of claim 1, 4, 6 or 7 , the expression cassette of claim 8 , the vector of claim 9 or 10, or the virion of claim 11 . 10. A composition for use in a method for producing an expression product comprising the synthetic CNS-specific expression cassette of claim 8 , the method comprising providing the synthetic CNS-specific expression cassette in a CNS cell and expressing an expression product present in the synthetic CNS-specific expression cassette , and optionally
a synthetic CNS-specific expression cassette comprising or consisting of SEQ ID NO:1 or SEQ ID NO:21 or a functional variant thereof, wherein when the expression cassette is provided by ICV injection, the expression product is broadly expressed in the brain, and optionally the CNS-specific expression cassette drives expression at a level of at least 100%, 150% or 200% compared to the activity of synapsin-1 (SEQ ID NO:14) in the brain; or
a synthetic CNS-specific expression cassette comprising or consisting of SEQ ID NO:8 or SEQ ID NO:26 or a functional variant thereof, wherein when administered by intravascular injection, the expression product is expressed in the midbrain, and optionally, the expression product is expressed in dopaminergic neurons;
Composition . 12. A composition for use in a method for expressing a therapeutic transgene in a CNS cell comprising the synthetic CNS-specific expression cassette of claim 8, the vector of claim 9 or 10, or the virion of claim 11 , the method comprising the step of introducing the synthetic CNS-specific expression cassette , vector or virion into a CNS cell , optionally wherein the expression cassette, vector or virion is introduced by intravenous injection, optionally wherein the injection is in one of the cephalic, median or basilic veins . A synthetic CNS-specific expression cassette according to claim 8, a vector according to claim 9 or 10, a virion according to claim 11 or a pharmaceutical composition according to claim 12 for use in a method of therapy of a subject, preferably a human, in need thereof,
The method comprises:
- administering to a subject said expression cassette , vector , virion or pharmaceutical composition comprising a sequence encoding a therapeutic product operably linked to a promoter according to claim 1, 4, 6 or 7 ; and
- expressing a therapeutic amount of a therapeutic product in the CNS of said subject;
An expression cassette, a vector, a virion or a pharmaceutical composition . 10. A composition for use in a method for expressing an expression product in a dopaminergic neuron comprising the synthetic CNS-specific expression cassette of claim 8, the method comprising introducing the synthetic CNS-specific expression cassette into a dopaminergic neuron by intravascular injection, the CNS-specific expression cassette comprising SEQ ID NO :8 or SEQ ID NO:26 or a functional variant thereof .