JPWO2020072354A5

JPWO2020072354A5 -

Info

Publication number: JPWO2020072354A5
Application number: JP2021518190A
Authority: JP
Publication date: 2022-10-06

Claims

An adeno-associated virus (AAV) having an AAVhu68 capsid and a vector genome comprising the GLB1 gene encoding human β-galactosidase under the control of regulatory sequences directing the expression of human β-galactosidase in target human cells.

said human β-galactosidase comprises a signal peptide and a mature β-galactosidase having an amino acid sequence of amino acids 24-677 of SEQ ID NO:4 ;
Optionally, the AAV of claim 1, wherein said signal peptide has the amino acid sequence of amino acids 1-23 of SEQ ID NO:4 .

Claim 1 or claim , wherein said GLB1 gene has the sequence of SEQ ID NO:8, or a sequence at least 95 % identical to SEQ ID NO:8, encoding said mature β-galactosidase from amino acids 24 to 677 of SEQ ID NO:4. Item 2. The AAV of item 2.

4. The AAV of claim 3, wherein the human β-galactosidase-encoding GLB1 gene encodes the amino acid sequence of SEQ ID NO:4.

3. The AAV of claim 1 or 2 , wherein said regulatory sequence comprises the human ubiquitin C (UbC) promoter.

wherein the vector genome is
(i) a 5' inverted terminal repeat (ITR);
(ii) the human ubiquitin C (UbC) promoter;
(iii) a chimeric intron;
(iv) the GLB1 gene encoding the native human signal peptide and human β-galactosidase;
(v) SV40 PolyA;
(vi) and 3' AAV ITRs
The AAV of claim 1, comprising:

said AAVhu68 capsid is produced from a sequence encoding the nucleic acid sequence of SEQ ID NO: 1 or the predicted amino acid sequence of SEQ ID NO: 2;
vp1 protein produced by expression from a nucleic acid sequence encoding the predicted amino acid sequence 1-736 of SEQ ID NO:2, the vp1 protein produced from SEQ ID NO:1, or the predicted amino acid sequence of 1-736 of SEQ ID NO:2 a heterogeneous population of AAVhu68 vp1 proteins selected from vp1 proteins produced from a nucleic acid sequence that is at least 70% identical to SEQ ID NO: 1 encoding amino acid sequence;
a vp2 protein produced by expression from a nucleic acid sequence encoding the predicted amino acid sequence of at least about amino acids 138-736 of SEQ ID NO:2, a vp2 protein produced from a sequence comprising at least nucleotides 412-2211 of SEQ ID NO:1; or an AAVhu68 vp2 protein selected from a vp2 protein produced from a nucleic acid sequence that is at least 70% identical to at least nucleotides 412-2211 of SEQ ID NO:1 that encodes the predicted amino acid sequence of at least about amino acids 138-736 of SEQ ID NO:2 a heterogeneous population of
a vp3 protein produced by expression from a nucleic acid sequence encoding the predicted amino acid sequence of at least about amino acids 203-736 of SEQ ID NO:2, a vp3 protein produced from a sequence comprising at least nucleotides 607-2211 of SEQ ID NO:1; or an AAVhu68 vp3 protein selected from a vp3 protein produced from a nucleic acid sequence that is at least 70% identical to at least nucleotides 607-2211 of SEQ ID NO:1 that encodes the predicted amino acid sequence of at least about amino acids 203-736 of SEQ ID NO:2 AAV according to any one of claims 1 to 5 , comprising a heterogeneous population of

An aqueous pharmaceutical composition comprising a formulation buffer and the AAV of any one of claims 1-5 .

The formulation buffer is
buffered saline and artificial cerebrospinal fluid comprising one or more of sodium, calcium, magnesium, potassium, or mixtures thereof;
a surfactant ;
9. The pharmaceutical composition of claim 8, wherein optionally said surfactant is present at 0.0005% w/w to about 0.001% w/w of said pharmaceutical composition.

10. The pharmaceutical composition of claim 8 or claim 9 , wherein said composition has a pH in the range of 7.5-7.8, or 6.2-7.7, or about 7.

An AAV according to any one of claims 1 to 5 or a pharmaceutical composition according to any one of claims 8 to 10 for use in the treatment of G M1 gangliosidosis.

wherein said AAV or said pharmaceutical composition comprises
(a) is it suitable for administration to a patient via intracisternal infusion (ICM);
(b) is it suitable for administration to patients with GM1 gangliosidosis;
(c) is suitable for administration to patients who are 18 months of age or younger or 18 months to 3 years of age; and/or
(d) ameliorating the symptoms of GM1 gangliosidosis or ameliorating the neurological symptoms of GM1 gangliosidosis, suitable for administration to a patient in need thereof, optionally said amelioration of GM1 gangliosidosis comprises increased life expectancy, reduced need for feeding tubes, reduced incidence and frequency of seizures, reduced progression to neurocognitive decline, and/or improved neurocognitive development ,
An AAV according to any one of claims 1 to 5 or a pharmaceutical composition according to any one of claims 8 to 10, for use according to claim 11.

11. The AAV or the pharmaceutical composition is administered into the cisterna magna via intracisternal infusion (ICM), optionally via CT-guided suboccipital infusion into the cisterna magna. An AAV according to any one of claims 1-5 or a pharmaceutical composition according to any one of claims 8-10 , for a use according to claims 11 or 12 .

AAV according to any one of claims 1 to 5 , for use according to any one of claims 11 to 13, wherein said AAV or said pharmaceutical composition is administered in a single dose, Or the pharmaceutical composition according to any one of claims 8-10 .

Any of claims 1 to 5, for use according to any one of claims 11 to 14, wherein said patient with GM1 gangliosidosis is 18 months or younger or between 18 months and 3 years of age. AAV according to one or a pharmaceutical composition according to any one of claims 8-10.

Administration of said AAV or composition ameliorates the symptoms of GM1 gangliosidosis or ameliorates the neurological symptoms of GM1 gangliosidosis, and optionally following treatment, the patient has an increased life expectancy , reduced need for a feeding tube, reduced incidence and frequency of seizures, reduced progression to neurocognitive decline, and/or improved neurocognitive development. An AAV according to any one of claims 1 to 5, or a pharmaceutical composition according to any one of claims 8 to 10, for the use according to one claim.

wherein said AAV is at a dose of 2×10 ¹² GC per patient to 3×10 ¹⁴ GC per patient, or at a dose of 8×10 ¹² genome copies (GC) per patient to 3×10 ¹⁴ GC per patient, optionally administered at a dose of 2 x ¹⁰ GC per patient to 3 x ¹⁰ GC per patient, 8 x ¹⁰ GC per patient to 3 x ¹⁰ GC per patient, or about 9 x ¹⁰ GC per patient; An AAV according to any one of claims 1-5 or a pharmaceutical composition according to any one of claims 8-10 , for a use according to any one of claims 11-17.

said AAV at a dose of 1× ^{10 10} GC/g brain mass to 3.4×10 ¹¹ GC/g brain mass, optionally 3.4×10 ¹⁰ GC/g brain mass to 3.4×10 ¹¹ GC/g brain mass, 1 .0×10 ¹¹ GC/g brain mass˜3 . Any of claims 1 to 5 , for use according to any of claims 11 to 17 , administered at a dose of 4×10 ¹¹ GC/g brain mass, or about 1.1×10 ¹¹ GC/g brain mass. AAV according to one or a pharmaceutical composition according to any one of claims 8-10 .