U.S. patent application number 16/184466 was filed with the patent office on 2019-02-28 for aadc polynucleotides for the treatment of parkinson's disease.
The applicant listed for this patent is VOYAGER THERAPEUTICS, INC.. Invention is credited to Bernard Ravina, Maria Scheel.
Application Number | 20190060425 16/184466 |
Document ID | / |
Family ID | 64659937 |
Filed Date | 2019-02-28 |
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United States Patent
Application |
20190060425 |
Kind Code |
A1 |
Scheel; Maria ; et
al. |
February 28, 2019 |
AADC POLYNUCLEOTIDES FOR THE TREATMENT OF PARKINSON'S DISEASE
Abstract
The disclosure relates to compositions and methods for the
preparation, manufacture and therapeutic use of polynucleotides
encoding AADC for the treatment of Parkinson's Disease.
Inventors: |
Scheel; Maria; (Cambridge,
MA) ; Ravina; Bernard; (Newton, MA) |
|
Applicant: |
Name |
City |
State |
Country |
Type |
VOYAGER THERAPEUTICS, INC. |
CAMBRIDGE |
MA |
US |
|
|
Family ID: |
64659937 |
Appl. No.: |
16/184466 |
Filed: |
November 8, 2018 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
|
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PCT/US2018/037437 |
Jun 14, 2018 |
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16184466 |
|
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62520084 |
Jun 15, 2017 |
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62554155 |
Sep 5, 2017 |
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Current U.S.
Class: |
1/1 |
Current CPC
Class: |
A61K 48/0066 20130101;
A61K 38/51 20130101; A61P 25/16 20180101; A61K 48/0075 20130101;
C12N 15/85 20130101; C12Y 401/01028 20130101; C12N 15/86 20130101;
C12N 2750/14143 20130101; A61K 48/005 20130101; C12N 15/62
20130101; C12N 9/88 20130101; A61K 35/761 20130101 |
International
Class: |
A61K 38/51 20060101
A61K038/51; C12N 15/85 20060101 C12N015/85; C12N 15/62 20060101
C12N015/62; A61P 25/16 20060101 A61P025/16; C12N 9/88 20060101
C12N009/88; A61K 48/00 20060101 A61K048/00; A61K 35/761 20060101
A61K035/761 |
Claims
1. An aromatic L-amino acid decarboxylase (AADC) polynucleotide
comprising an AADC sequence region which has at least 95% identity
to SEQ ID NO: 979.
2. The AADC polynucleotide of claim 1, wherein the AADC sequence
region comprises a promoter region, an enhancer region, a multiple
cloning site (MCS) region and a polyadenylation (poly(A)) signal
region.
3. The AADC polynucleotide of claim 2, wherein the AADC sequence
region comprises at least one 5' inverted terminal repeat (ITR)
region and one 3' ITR region.
4. The AADC polynucleotide of claim 3, wherein one or more of the
5' ITRs are located 5' to the MCS region and one or more of the 3'
ITRs are located 3' to the poly(A) signal.
5. The AADC polynucleotide of claim 4, wherein the AADC sequence
region comprises a first exon region, a first intron region, a
second intron region and a second exon region.
6. The AADC polynucleotide of claim 5, wherein the enhancer region
and the promoter region are derived from CMV.
7. The AADC polynucleotide of claim 6, wherein the first exon
region is immediate-early 1 (ie1) exon 1 or fragments thereof, the
first intron region is ie1 intron 1 or fragments thereof, the
second intron region is human beta-globin (hBglobin) intron 2 or
fragments thereof and the second exon region is hBglobin exon 3 or
fragments thereof.
8. The AADC polynucleotide of claim 7, wherein the poly(A) signal
is derived from human growth hormone.
9. The AADC polynucleotide of claim 8, wherein the AADC sequence
region consists of 3520-3530 nucleotides from the 5' end of the 5'
ITR to the 3' end of the 3' ITR.
10. The AADC polynucleotide of claim 8, wherein the AADC sequence
region consists of 3526 nucleotides from the 5' end of the 5' ITR
to the 3' end of the 3' ITR.
11. A recombinant adeno-associated virus (rAAV) comprising the AADC
polynucleotide of claim 8.
12. The rAAV virus of claim 11, wherein the capsid serotype of the
rAAV is AAV2.
13. The AADC polynucleotide of claim 3, wherein the AADC sequence
region has at least 99% identity to SEQ ID NO: 979.
14. The AADC polynucleotide of claim 13, wherein the AADC sequence
region consists of 3526 nucleotides from the 5' end of the 5' ITR
to the 3' end of the 3' ITR.
15. A recombinant adeno-associated virus (rAAV) comprising the AADC
polynucleotide of claim 13.
16. The rAAV virus of claim 15, wherein the capsid serotype of the
rAAV is AAV2.
17. A pharmaceutical composition comprising an adeno-associated
virus (AAV) particle, wherein the AAV particle comprises an AAV
capsid and a vector genome, and wherein the vector genome comprises
at least one AADC sequence region with at least 95% identity to SEQ
ID NO: 979.
18. The pharmaceutical composition of claim 17, wherein the AADC
sequence region has at least 99% identity SEQ ID NO: 979.
19. The pharmaceutical composition of claim 18, wherein the AAV
capsid serotype is AAV2.
20. The pharmaceutical composition of claim 19, wherein at least
70% of the AAV particles contain a vector genome.
Description
CROSS REFERENCE TO RELATED APPLICATIONS
[0001] The present application is a continuation application which
claims priority to PCT/US2018/037437, filed Jun. 14, 2018, entitled
AADC POLYNUCLEOTIDES FOR THE TREATMENT OF PARKINSON'S DISEASE;
which claims priority to U.S. provisional application U.S.
62/554,155, filed Sep. 5, 2017, entitled AADC POLYNUCLEOTIDES FOR
THE TREATMENT OF PARKINSON'S DISEASE, and to U.S. provisional
application U.S. 62/520,084, filed Jun. 15, 2017, entitled AADC
POLYNUCLEOTIDES FOR THE TREATMENT OF PARKINSON'S DISEASE; the
contents of each being incorporated by reference herein in their
entirety.
REFERENCE TO SEQUENCE LISTING
[0002] The present application is being filed along with a Sequence
Listing in electronic format. The Sequence Listing file, entitled
20571042USCONSEQLST, was created on Nov. 8, 2018, and is 6,421,705
bytes in size. The information in electronic format of the Sequence
Listing is incorporated herein by reference in its entirety.
FIELD OF THE INVENTION
[0003] The invention relates to compositions, particularly nucleic
acid molecules, e.g., polynucleotides encoding AADC, for use in the
treatment of Parkinson's disease. In some embodiments such AADC
polynucleotides may be encoded by or within recombinant
adeno-associated viruses (AAVs).
BACKGROUND
[0004] Aromatic L-amino acid decarboxylase (AADC) is a homodimeric
pyridoxal phosphate-dependent enzyme responsible for the synthesis
of dopamine and serotonin. The encoded protein catalyzes the
decarboxylation of L-3,4-dihydroxyphenylalanine (L-DOPA or
levodopa) to dopamine; L-5-hydroxytryptophan to serotonin; and
L-tryptophan to tryptamine. Defects in this gene are the cause of
aromatic L-amino-acid decarboxylase deficiency (AADCD), which is an
inborn error in neurotransmitter metabolism leading to combined
serotonin and catecholamine deficiency that results in severe motor
and autonomic dysfunctions.
[0005] Parkinson's Disease (PD) is a progressive neurodegenerative
disease of the central nervous system (CNS) producing sensory and
motor symptoms. Dopamine replacement (i.e., levodopa) has been the
standard pharmacotherapy for motor impairment in PD. However, the
benefit of dopamine therapy becomes less marked over time, due, in
part, to the progressive death of dopamine-generating cells and
corresponding loss of AADC activity. Furthermore, systemic
administration of high-dose dopamine is complicated by side
effects, such as fluctuations in motor performance, dyskinesias,
and hallucinations, resulting from dopaminergic stimulation of the
mesolimbic system. One strategy to restore dopaminergic function
and minimize side effects is the use of gene therapy to deliver
AADC directly to a targeted region of the CNS.
[0006] The adeno-associated virus (AAV) has emerged as an
attractive vector for gene therapy due to its long-term gene
expression, the inability to autonomously replicate without a
helpervirus, the ability to transduce dividing and non-diving
cells, and the lack of pathogenicity from wild-type infections (See
e.g., Hadaczek et al. Mol. Ther. 18(8), 1458-1461, August 2010).
AAV is a helper-dependent DNA parvovirus which belongs to the genus
Dependovirus.
[0007] The present disclosure provides such improved nucleic acid
constructs, e.g., polynucleotides, for use with AAV-derived vectors
comprising dopa carboxylase ("DDC") gene sequence which encodes a
full-length AADC protein for the purpose of gene therapy in the
treatment of Parkinson's Disease.
[0008] The nucleic acid constructs described herein comprise at
least a 5'-ITR and a 3'-ITR, each or both of which may be derived
from an AAV, positioned about a DDC gene sequence, as well as
additional components required for gene expression and clone
selection.
SUMMARY
[0009] Described herein are compositions, methods, processes, kits
and devices for the design, preparation, manufacture and/or
formulation of AADC polynucleotides.
[0010] In some embodiments such AADC polynucleotides may be encoded
by or contained within plasmids or vectors or recombinant
adeno-associated viruses (AAV).
[0011] The details of various embodiments of the disclosure are set
forth in the description below. Other features, objects, and
advantages of the disclosure will be apparent from the description
and from the claims.
BRIEF DESCRIPTION OF THE DRAWINGS
[0012] The foregoing and other objects, features and advantages
will be apparent from the following description of particular
embodiments of the disclosure, as illustrated in the accompanying
drawings. The drawings are not necessarily to scale, emphasis
instead being placed upon illustrating the principles of various
embodiments of the disclosure.
[0013] FIG. 1 is a schematic of a viral genome of the
disclosure.
DETAILED DESCRIPTION
I. Compositions
Adeno-Associated Viruses (AAVs) and AAV Particles
[0014] Viruses of the Parvoviridae family are small non-enveloped
icosahedral capsid viruses characterized by a single stranded DNA
genome. Parvoviridae family viruses consist of two subfamilies:
Parvovirinae, which infect vertebrates, and Densovirinae, which
infect invertebrates. Due to its relatively simple structure,
easily manipulated using standard molecular biology techniques,
this virus family is useful as a biological tool. The genome of the
virus may be modified to contain a minimum of components for the
assembly of a functional recombinant virus, or viral particle,
which is loaded with or engineered to express or deliver a desired
payload, which may be delivered to a target cell, tissue, organ, or
organism.
[0015] The parvoviruses and other members of the Parvoviridae
family are generally described in Kenneth I. Berns, "Parvoviridae:
The Viruses and Their Replication," Chapter 69 in FIELDS VIROLOGY
(3d Ed. 1996), the contents of which are incorporated by reference
in their entirety.
[0016] The Parvoviridae family comprises the Dependovirus genus
which includes adeno-associated viruses (AAV) capable of
replication in vertebrate hosts including, but not limited to,
human, primate, bovine, canine, equine, and ovine species.
[0017] The vector genome is a linear, single-stranded DNA (ssDNA)
molecule approximately 5,000 nucleotides (nt) in length. The AAV
viral genome can comprise a payload region and at least one
inverted terminal repeat (ITR) or ITR region. ITRs traditionally
flank the coding nucleotide sequences for the non-structural
proteins (encoded by Rep genes) and the structural proteins
(encoded by capsid genes or Cap genes). While not wishing to be
bound by theory, an AAV viral genome typically comprises two ITR
sequences. The vector genome comprises a characteristic T-shaped
hairpin structure defined by the self-complementary terminal 145 nt
of the 5' and 3' ends of the ssDNA which form an energetically
stable double stranded region. The double stranded hairpin
structures comprise multiple functions including, but not limited
to, acting as an origin for DNA replication by functioning as
primers for the endogenous DNA polymerase complex of the host viral
replication cell.
[0018] In addition to the encoded heterologous payload, AAV
particles may comprise the viral genome, in whole or in part, of
any naturally occurring and/or recombinant AAV serotype nucleotide
sequence or variant. AAV variants may have sequences of significant
homology at the nucleic acid (genome or capsid) and amino acid
levels (capsids), to produce constructs which are generally
physical and functional equivalents, replicate by similar
mechanisms, and assemble by similar mechanisms. Chiorini et al., J.
Vir. 71: 6823-33 (1997); Srivastava et al., J. Vir. 45:555-64
(1983); Chiorini et al., J. Vir. 73:1309-1319 (1999); Rutledge et
al., J. Vir. 72:309-319 (1998); and Wu et al., J. Vir. 74: 8635-47
(2000), the contents of each of which are incorporated herein by
reference in their entirety.
[0019] In one embodiment, AAV particles of the present disclosure
are recombinant AAV particles which are replication defective,
lacking sequences encoding functional Rep and Cap proteins within
their viral genome. These defective AAV particles may lack most or
all parental coding sequences and essentially carry only one or two
AAV ITR sequences and the nucleic acid of interest for delivery to
a cell, a tissue, an organ or an organism.
[0020] In one embodiment, the viral genome of the AAV particles of
the present disclosure comprise at least one control element which
provides for the replication, transcription and translation of a
coding sequence encoded therein. Not all of the control elements
need always be present as long as the coding sequence is capable of
being replicated, transcribed and/or translated in an appropriate
host cell. Non-limiting examples of expression control elements
include sequences for transcription initiation and/or termination,
promoter and/or enhancer sequences, efficient RNA processing
signals such as splicing and polyadenylation signals, sequences
that stabilize cytoplasmic mRNA, sequences that enhance translation
efficacy (e.g., Kozak consensus sequence), sequences that enhance
protein stability, and/or sequences that enhance protein processing
and/or secretion.
[0021] According to the present disclosure, AAV particles for use
in therapeutics and/or diagnostics comprise a virus that has been
distilled or reduced to the minimum components necessary for
transduction of a nucleic acid payload or cargo of interest. In
this manner, AAV particles are engineered as vehicles for specific
delivery while lacking the deleterious replication and/or
integration features found in wild-type viruses.
[0022] AAV particles of the present disclosure may be produced
recombinantly and may be based on adeno-associated virus (AAV)
parent or reference sequences. As used herein, a "vector" is any
molecule or moiety which transports, transduces or otherwise acts
as a carrier of a heterologous molecule such as the nucleic acids
described herein.
[0023] In addition to single stranded AAV particles (e.g., ssAAVs),
the present disclosure also provides for self-complementary AAV
(scAAVs) particles. scAAV particles contain DNA strands which
anneal together to form double stranded DNA. By skipping second
strand synthesis, scAAVs allow for rapid expression in the
cell.
[0024] In one embodiment, the AAV particle of the present
disclosure is an scAAV.
[0025] In one embodiment, the AAV particle of the present
disclosure is an ssAAV.
[0026] Methods for producing and/or modifying AAV particles are
disclosed in the art such as pseudotyped AAV particles (PCT Patent
Publication Nos. WO200028004; WO200123001; WO2004112727; WO
2005005610 and WO 2005072364, the content of each of which is
incorporated herein by reference in its entirety).
[0027] AAV particles may be modified to enhance the efficiency of
delivery. Such modified AAV particles can be packaged efficiently
and be used to successfully infect the target cells at high
frequency and with minimal toxicity. In some embodiments the
capsids of the AAV particles are engineered according to the
methods described in US Publication Number US 20130195801, the
contents of which are incorporated herein by reference in their
entirety.
[0028] In one embodiment, the AAV particles comprising a payload
region encoding the polypeptides of the disclosure may be
introduced into mammalian cells.
AAV Serotypes
[0029] AAV particles of the present disclosure may comprise or be
derived from any natural or recombinant AAV serotype. According to
the present disclosure, the AAV particles may utilize or be based
on a serotype selected from any of the following PHP.B, PHP.A,
AAV1, AAV2, AAV2G9, AAV3, AAV3a, AAV3b, AAV3-3, AAV4, AAV4-4, AAV5,
AAV6, AAV6.1, AAV6.2, AAV6.1.2, AAV7, AAV7.2, AAV8, AAV9, AAV9.11,
AAV9.13, AAV9.16, AAV9.24, AAV9.45, AAV9.47, AAV9.61, AAV9.68,
AAV9.84, AAV9.9, AAV10, AAV11, AAV12, AAV16.3, AAV24.1, AAV27.3,
AAV42.12, AAV42-lb, AAV42-2, AAV42-3a, AAV42-3b, AAV42-4, AAV42-5a,
AAV42-5b, AAV42-6b, AAV42-8, AAV42-10, AAV42-11, AAV42-12,
AAV42-13, AAV42-15, AAV42-aa, AAV43-1, AAV43-12, AAV43-20,
AAV43-21, AAV43-23, AAV43-25, AAV43-5, AAV44.1, AAV44.2, AAV44.5,
AAV223.1, AAV223.2, AAV223.4, AAV223.5, AAV223.6, AAV223.7,
AAV1-7/rh.48, AAV1-8/rh.49, AAV2-15/rh.62, AAV2-3/rh.61,
AAV2-4/rh.50, AAV2-5/rh.51, AAV3.1/hu.6, AAV3.1/hu.9, AAV3-9/rh.52,
AAV3-11/rh. 53, AAV4-8/r11.64, AAV4-9/rh. 54, AAV4-19/rh.55,
AAV5-3/rh.57, AAV5-22/rh.58, AAV7.3/hu.7, AAV16.8/hu.10,
AAV16.12/hu.11, AAV29.3/bb.1, AAV29.5/bb.2, AAV106.1/hu.37,
AAV114.3/hu.40, AAV127.2/hu.41, AAV127.5/hu.42, AAV128.3/hu.44,
AAV130.4/hu.48, AAV145.1/hu.53, AAV145.5/hu.54, AAV145.6/hu.55,
AAV161.10/hu.60, AAV161.6/hu.61, AAV33.12/hu.17, AAV33.4/hu.15,
AAV33.8/hu. 16, AAV52/hu. 19, AAV52.1/hu.20, AAV58.2/hu.25,
AAVA3.3, AAVA3.4, AAVA3.5, AAVA3.7, AAVC1, AAVC2, AAVC5, AAV-DJ,
AAV-DJ8, AAVF3, AAVF5, AAVH2, AAVrh.72, AAVhu.8, AAVrh.68,
AAVrh.70, AAVpi.1, AAVpi.3, AAVpi.2, AAVrh.60, AAVrh.44, AAVrh.65,
AAVrh.55, AAVrh.47, AAVrh.69, AAVrh.45, AAVrh.59, AAVhu.12, AAVH6,
AAVLK03, AAVH-1/hu. 1, AAVH-5/hu.3, AAVLG-10/rh.40, AAVLG-4/rh. 38,
AAVLG-9/hu. 39, AAVN721-8/rh.43, AAVCh.5, AAVCh.5R1, AAVcy.2,
AAVcy.3, AAVcy.4, AAVcy.5, AAVCy.5R1, AAVCy.5R2, AAVCy.5R3,
AAVCy.5R4, AAVcy.6, AAVhu.1, AAVhu.2, AAVhu.3, AAVhu.4, AAVhu.5,
AAVhu.6, AAVhu.7, AAVhu.9, AAVhu.10, AAVhu.11, AAVhu.13, AAVhu.15,
AAVhu.16, AAVhu.17, AAVhu.18, AAVhu.20, AAVhu.21, AAVhu.22,
AAVhu.23.2, AAVhu.24, AAVhu.25, AAVhu.27, AAVhu.28, AAVhu.29,
AAVhu.29R, AAVhu.31, AAVhu.32, AAVhu.34, AAVhu.35, AAVhu.37,
AAVhu.39, AAVhu.40, AAVhu.41, AAVhu.42, AAVhu.43, AAVhu.44,
AAVhu.44R1, AAVhu.44R2, AAVhu.44R3, AAVhu.45, AAVhu.46, AAVhu.47,
AAVhu.48, AAVhu.48R1, AAVhu.48R2, AAVhu.48R3, AAVhu.49, AAVhu.51,
AAVhu.52, AAVhu.54, AAVhu.55, AAVhu.56, AAVhu.57, AAVhu.58,
AAVhu.60, AAVhu.61, AAVhu.63, AAVhu.64, AAVhu.66, AAVhu.67,
AAVhu.14/9, AAVhu.t 19, AAVrh.2, AAVrh.2R, AAVrh.8, AAVrh.8R,
AAVrh.10, AAVrh.12, AAVrh.13, AAVrh.13R, AAVrh.14, AAVrh.17,
AAVrh.18, AAVrh.19, AAVrh.20, AAVrh.21, AAVrh.22, AAVrh.23,
AAVrh.24, AAVrh.25, AAVrh.31, AAVrh.32, AAVrh.33, AAVrh.34,
AAVrh.35, AAVrh.36, AAVrh.37, AAVrh.37R2, AAVrh.38, AAVrh.39,
AAVrh.40, AAVrh.46, AAVrh.48, AAVrh.48.1, AAVrh.48.1.2, AAVrh.48.2,
AAVrh.49, AAVrh.51, AAVrh.52, AAVrh.53, AAVrh.54, AAVrh.56,
AAVrh.57, AAVrh.58, AAVrh.61, AAVrh.64, AAVrh.64R1, AAVrh.64R2,
AAVrh.67, AAVrh.73, AAVrh.74, AAVrh8R, AAVrh8R A586R mutant,
AAVrh8R R533A mutant, AAAV, BAAV, caprine AAV, bovine AAV, ovine
AAV, AAVhE1.1, AAVhEr1.5, AAVhER1.14, AAVhEr1.8, AAVhEr1.16,
AAVhEr1.18, AAVhEr1.35, AAVhEr1.7, AAVhEr1.36, AAVhEr2.29,
AAVhEr2.4, AAVhEr2.16, AAVhEr2.30, AAVhEr2.31, AAVhEr2.36,
AAVhER1.23, AAVhEr3.1, AAV2.5T, AAV-PAEC, AAV-LK01, AAV-LK02,
AAV-LK03, AAV-LK04, AAV-LK05, AAV-LK06, AAV-LK07, AAV-LK08,
AAV-LK09, AAV-LK10, AAV-LK11, AAV-LK12, AAV-LK13, AAV-LK14,
AAV-LK15, AAV-LK16, AAV-LK17, AAV-LK18, AAV-LK19, AAV-PAEC2,
AAV-PAEC4, AAV-PAEC6, AAV-PAEC7, AAV-PAEC8, AAV-PAEC11, AAV-PAEC12,
AAV-2-pre-miRNA-101, AAV-8h, AAV-8b, AAV-h, AAV-b, AAV SM 10-2, AAV
Shuffle 100-1, AAV Shuffle 100-3, AAV Shuffle 100-7, AAV Shuffle
10-2, AAV Shuffle 10-6, AAV Shuffle 10-8, AAV Shuffle 100-2, AAV SM
10-1, AAV SM 10-8, AAV SM 100-3, AAV SM 100-10, BNP61 AAV, BNP62
AAV, BNP63 AAV, AAVrh.50, AAVrh.43, AAVrh.62, AAVrh.48, AAVhu. 19,
AAVhu. 11, AAVhu.53, AAV4-8/rh.64, AAVLG-9/hu.39, AAV54.5/hu.23,
AAV54.2/hu.22, AAV54.7/hu.24, AAV54.1/hu.21, AAV54.4R/hu.27,
AAV46.2/hu.28, AAV46.6/hu.29, AAV128.1/hu.43, true type AAV
(ttAAV), UPENN AAV 10, Japanese AAV 10 serotypes, AAV CBr-7.1, AAV
CBr-7.10, AAV CBr-7.2, AAV CBr-7.3, AAV CBr-7.4, AAV CBr-7.5, AAV
CBr-7.7, AAV CBr-7.8, AAV CBr-B7.3, AAV CBr-B7.4, AAV CBr-E1, AAV
CBr-E2, AAV CBr-E3, AAV CBr-E4, AAV CBr-E5, AAV CBr-e5, AAV CBr-E6,
AAV CBr-E7, AAV CBr-E8, AAV CHt-1, AAV CHt-2, AAV CHt-3, AAV
CHt-6.1, AAV CHt-6.10, AAV CHt-6.5, AAV CHt-6.6, AAV CHt-6.7, AAV
CHt-6.8, AAV CHt-P1, AAV CHt-P2, AAV CHt-P5, AAV CHt-P6, AAV
CHt-P8, AAV CHt-P9, AAV CKd-1, AAV CKd-10, AAV CKd-2, AAV CKd-3,
AAV CKd-4, AAV CKd-6, AAV CKd-7, AAV CKd-8, AAV CKd-B1, AAV CKd-B2,
AAV CKd-B3, AAV CKd-B4, AAV CKd-B5, AAV CKd-B6, AAV CKd-B7, AAV
CKd-B8, AAV CKd-H1, AAV CKd-H2, AAV CKd-H3, AAV CKd-H4, AAV CKd-H5,
AAV CKd-H6, AAV CKd-N3, AAV CKd-N4, AAV CKd-N9, AAV CLg-F1, AAV
CLg-F2, AAV CLg-F3, AAV CLg-F4, AAV CLg-F5, AAV CLg-F6, AAV CLg-F7,
AAV CLg-F8, AAV CLv-1, AAV CLv1-1, AAV Clv1-10, AAV CLv1-2, AAV
CLv-12, AAV CLv1-3, AAV CLv-13, AAV CLv1-4, AAV Clv1-7, AAV Clv1-8,
AAV Clv1-9, AAV CLv-2, AAV CLv-3, AAV CLv-4, AAV CLv-6, AAV CLv-8,
AAV CLv-D1, AAV CLv-D2, AAV CLv-D3, AAV CLv-D4, AAV CLv-D5, AAV
CLv-D6, AAV CLv-D7, AAV CLv-D8, AAV CLv-E1, AAV CLv-K1, AAV CLv-K3,
AAV CLv-K6, AAV CLv-L4, AAV CLv-L5, AAV CLv-L6, AAV CLv-M1, AAV
CLv-M11, AAV CLv-M2, AAV CLv-M5, AAV CLv-M6, AAV CLv-M7, AAV
CLv-M8, AAV CLv-M9, AAV CLv-R1, AAV CLv-R2, AAV CLv-R3, AAV CLv-R4,
AAV CLv-R5, AAV CLv-R6, AAV CLv-R7, AAV CLv-R8, AAV CLv-R9, AAV
CSp-1, AAV CSp-10, AAV CSp-11, AAV CSp-2, AAV CSp-3, AAV CSp-4, AAV
CSp-6, AAV CSp-7, AAV CSp-8, AAV CSp-8.10, AAV CSp-8.2, AAV
CSp-8.4, AAV CSp-8.5, AAV CSp-8.6, AAV CSp-8.7, AAV CSp-8.8, AAV
CSp-8.9, AAV CSp-9, AAV.hu.48R3, AAV.VR-355, AAV3B, AAV4, AAV5,
AAVF1/HSC1, AAVF11/HSC11, AAVF12/HSC12, AAVF13/HSC13, AAVF14/HSC14,
AAVF15/HSC15, AAVF16/HSC16, AAVF17/HSC17, AAVF2/HSC2, AAVF3/HSC3,
AAVF4/HSC4, AAVF5/HSC5, AAVF6/HSC6, AAVF7/HSC7, AAVF8/HSC8,
AAVF9/HSC9, PHP.B (AAV-PHP.B), PHP.A (AAV.PHP.A), G2B-26, G2B-13,
TH1.1-32, TH1.1-35, AAVPHP.B2, AAVPHP.B3, AAVPHP.N/PHP.B-DGT,
AAVPHP.B-EST, AAVPHP.B-GGT, AAVPHP.B-ATP, AAVPHP.B-ATT-T,
AAVPHP.B-DGT-T, AAVPHP.B-GGT-T, AAVPHP.B-SGS, AAVPHP.B-AQP,
AAVPHP.B-QQP, AAVPHP.B-SNP(3), AAVPHP.B-SNP, AAVPHP.B-QGT,
AAVPHP.B-NQT, AAVPHP.B-EGS, AAVPHP.B-SGN, AAVPHP.B-EGT,
AAVPHP.B-DST, AAVPHP.B-DST, AAVPHP.B-STP, AAVPHP.B-PQP,
AAVPHP.B-SQP, AAVPHP.B-QLP, AAVPHP.B-TMP, AAVPHP.B-TTP,
AAVPHP.S/G2A12, AAVG2A15/G2A3, AAVG2B4, and/or AAVG2B5, and
variants thereof.
[0030] In some embodiments, the AAV serotype may be, or have, a
modification as described in United States Publication No. US
20160361439, the contents of which are herein incorporated by
reference in their entirety, such as but not limited to, Y252F,
Y272F, Y444F, Y500F, Y700F, Y704F, Y730F, Y275F, Y281F, Y508F,
Y576F, Y612G, Y673F, and Y720F of the wild-type AAV1, AAV2, AAV3,
AAV4, AAV5, AAV6, AAV7, AAV8, AAV9, AAV10, AAV11, AAV12, and
hybrids thereof.
[0031] In some embodiments, the AAV serotype may be, or have, a
mutation as described in U.S. Pat. No. 9,546,112, the contents of
which are herein incorporated by reference in their entirety, such
as, but not limited to, at least two, but not all the F129L, D418E,
K531E, L584F, V598A and H642N mutations in the sequence of AAV6
(SEQ ID NO:4 of U.S. Pat. No. 9,546,112), AAV1 (SEQ ID NO:6 of U.S.
Pat. No. 9,546,112), AAV2, AAV3, AAV4, AAV5, AAV7, AAV9, AAV10 or
AAV11 or derivatives thereof. In yet another embodiment, the AAV
serotype may be, or have, an AAV6 sequence comprising the K531E
mutation (SEQ ID NO:5 of U.S. Pat. No. 9,546,112).
[0032] In some embodiments, the AAV serotype may be, or have, a
mutation in the AAV1 sequence, as described in in United States
Publication No. US 20130224836, the contents of which are herein
incorporated by reference in their entirety, such as, but not
limited to, at least one of the surface-exposed tyrosine residues,
preferably, at positions 252, 273, 445, 701, 705 and 731 of AAV1
(SEQ ID NO: 2 of US 20130224836) substituted with another amino
acid, preferably with a phenylalanine residue. In one embodiment,
the AAV serotype may be, or have, a mutation in the AAV9 sequence,
such as, but not limited to, at least one of the surface-exposed
tyrosine residues, preferably, at positions 252, 272, 444, 500,
700, 704 and 730 of AAV2 (SEQ ID NO: 4 of US 20130224836)
substituted with another amino acid, preferably with a
phenylalanine residue. In one embodiment, the tyrosine residue at
position 446 of AAV9 (SEQ ID NO: 6 US 20130224836) is substituted
with a phenylalanine residue.
[0033] In some embodiments, the serotype may be AAV2 or a variant
thereof, as described in International Publication No.
WO2016130589, herein incorporated by reference in its entirety. The
amino acid sequence of AAV2 may comprise N587A, E548A, or N708A
mutations. In one embodiment, the amino acid sequence of any AAV
may comprise a V708K mutation.
[0034] In some embodiments, the AAV serotype may be, or have, a
sequence as described in United States Publication No.
US20030138772, the contents of which are herein incorporated by
reference in their entirety, such as, but not limited to, AAV1 (SEQ
ID NO: 6 and 64 of US20030138772), AAV2 (SEQ ID NO: 7 and 70 of
US20030138772), AAV3 (SEQ ID NO: 8 and 71 of US20030138772), AAV4
(SEQ ID NO: 63 of US20030138772), AAV5 (SEQ ID NO: 114 of
US20030138772), AAV6 (SEQ ID NO: 65 of US20030138772), AAV7 (SEQ ID
NO: 1-3 of US20030138772), AAV8 (SEQ ID NO: 4 and 95 of
US20030138772), AAV9 (SEQ ID NO: 5 and 100 of US20030138772), AAV10
(SEQ ID NO: 117 of US20030138772), AAV11 (SEQ ID NO: 118 of
US20030138772), AAV12 (SEQ ID NO: 119 of US20030138772), AAVrh10
(amino acids 1 to 738 of SEQ ID NO: 81 of US20030138772), AAV16.3
(US20030138772 SEQ ID NO: 10), AAV29.3/bb.1 (US20030138772 SEQ ID
NO: 11), AAV29.4 (US20030138772 SEQ ID NO: 12), AAV29.5/bb.2
(US20030138772 SEQ ID NO: 13), AAV1.3 (US20030138772 SEQ ID NO:
14), AAV13.3 (US20030138772 SEQ ID NO: 15), AAV24.1 (US20030138772
SEQ ID NO: 16), AAV27.3 (US20030138772 SEQ ID NO: 17), AAV7.2
(US20030138772 SEQ ID NO: 18), AAVC1 (US20030138772 SEQ ID NO: 19),
AAVC3 (US20030138772 SEQ ID NO: 20), AAVC5 (US20030138772 SEQ ID
NO: 21), AAVF1 (US20030138772 SEQ ID NO: 22), AAVF3 (US20030138772
SEQ ID NO: 23), AAVF5 (US20030138772 SEQ ID NO: 24), AAVH6
(US20030138772 SEQ ID NO: 25), AAVH2 (US20030138772 SEQ ID NO: 26),
AAV42-8 (US20030138772 SEQ ID NO: 27), AAV42-15 (US20030138772 SEQ
ID NO: 28), AAV42-5b (US20030138772 SEQ ID NO: 29), AAV42-1b
(US20030138772 SEQ ID NO: 30), AAV42-13 (US20030138772 SEQ ID NO:
31), AAV42-3a (US20030138772 SEQ ID NO: 32), AAV42-4 (US20030138772
SEQ ID NO: 33), AAV42-5a (US20030138772 SEQ ID NO: 34), AAV42-10
(US20030138772 SEQ ID NO: 35), AAV42-3b (US20030138772 SEQ ID NO:
36), AAV42-11 (US20030138772 SEQ ID NO: 37), AAV42-6b
(US20030138772 SEQ ID NO: 38), AAV43-1 (US20030138772 SEQ ID NO:
39), AAV43-5 (US20030138772 SEQ ID NO: 40), AAV43-12 (US20030138772
SEQ ID NO: 41), AAV43-20 (US20030138772 SEQ ID NO: 42), AAV43-21
(US20030138772 SEQ ID NO: 43), AAV43-23 (US20030138772 SEQ ID NO:
44), AAV43-25 (US20030138772 SEQ ID NO: 45), AAV44.1 (US20030138772
SEQ ID NO: 46), AAV44.5 (U.S. Pat. No. 2,003,013 8772 SEQ ID NO:
47), AAV223.1 (US20030138772 SEQ ID NO: 48), AAV223.2
(US20030138772 SEQ ID NO: 49), AAV223.4 (US20030138772 SEQ ID NO:
50), AAV223.5 (US20030138772 SEQ ID NO: 51), AAV223.6
(US20030138772 SEQ ID NO: 52), AAV223.7 (US20030138772 SEQ ID NO:
53), AAVA3.4 (US20030138772 SEQ ID NO: 54), AAVA3.5 (US20030138772
SEQ ID NO: 55), AAVA3.7 (US20030138772 SEQ ID NO: 56), AAVA3.3
(US20030138772 SEQ ID NO: 57), AAV42.12 (US20030138772 SEQ ID NO:
58), AAV44.2 (US20030138772 SEQ ID NO: 59), AAV42-2 (US20030138772
SEQ ID NO: 9), or variants thereof.
[0035] In some embodiments, the AAV serotype may be, or have, a
sequence as described in United States Publication No.
US20150159173, the contents of which are herein incorporated by
reference in their entirety, such as, but not limited to, AAV2 (SEQ
ID NO: 7 and 23 of US20150159173), rh20 (SEQ ID NO: 1 of
US20150159173), rh32/33 (SEQ ID NO: 2 of US20150159173), rh39 (SEQ
ID NO: 3, 20 and 36 of US20150159173), rh46 (SEQ ID NO: 4 and 22 of
US20150159173), rh73 (SEQ ID NO: 5 of US20150159173), rh74 (SEQ ID
NO: 6 of US20150159173), AAV6.1 (SEQ ID NO: 29 of US20150159173),
rh.8 (SEQ ID NO: 41 of US20150159173), rh.48.1 (SEQ ID NO: 44 of
US20150159173), hu.44 (SEQ ID NO: 45 of US20150159173), hu.29 (SEQ
ID NO: 42 of US20150159173), hu.48 (SEQ ID NO: 38 of
US20150159173), rh54 (SEQ ID NO: 49 of US20150159173), AAV2 (SEQ ID
NO: 7 of US20150159173), cy.5 (SEQ ID NO: 8 and 24 of
US20150159173), rh.10 (SEQ ID NO: 9 and 25 of US20150159173), rh.13
(SEQ ID NO: 10 and 26 of US20150159173), AAV1 (SEQ ID NO: 11 and 27
of US20150159173), AAV3 (SEQ ID NO: 12 and 28 of US20150159173),
AAV6 (SEQ ID NO: 13 and 29 of US20150159173), AAV7 (SEQ ID NO: 14
and 30 of US20150159173), AAV8 (SEQ ID NO: 15 and 31 of
US20150159173), hu.13 (SEQ ID NO: 16 and 32 of US20150159173),
hu.26 (SEQ ID NO: 17 and 33 of US20150159173), hu.37 (SEQ ID NO: 18
and 34 of US20150159173), hu.53 (SEQ ID NO: 19 and 35 of
US20150159173), rh.43 (SEQ ID NO: 21 and 37 of US20150159173), rh2
(SEQ ID NO: 39 of US20150159173), rh.37 (SEQ ID NO: 40 of
US20150159173), rh.64 (SEQ ID NO: 43 of US20150159173), rh.48 (SEQ
ID NO: 44 of US20150159173), ch.5 (SEQ ID NO 46 of US20150159173),
rh.67 (SEQ ID NO: 47 of US20150159173), rh.58 (SEQ ID NO: 48 of
US20150159173), or variants thereof including, but not limited to
Cy5R1, Cy5R2, Cy5R3, Cy5R4, rh.13R, rh.37R2, rh.2R, rh.8R, rh.48.1,
rh.48.2, rh.48.1.2, hu.44R1, hu.44R2, hu.44R3, hu.29R, ch.5R1,
rh64R1, rh64R2, AAV6.2, AAV6.1, AAV6.12, hu.48R1, hu.48R2, and
hu.48R3.
[0036] In some embodiments, the AAV serotype may be, or have, a
sequence as described in U.S. Pat. No. 7,198,951, the contents of
which are herein incorporated by reference in their entirety, such
as, but not limited to, AAV9 (SEQ ID NO: 1-3 of U.S. Pat. No.
7,198,951), AAV2 (SEQ ID NO: 4 of U.S. Pat. No. 7,198,951), AAV1
(SEQ ID NO: 5 of U.S. Pat. No. 7,198,951), AAV3 (SEQ ID NO: 6 of
U.S. Pat. No. 7,198,951), and AAV8 (SEQ ID NO: 7 of US7198951).
[0037] In some embodiments, the AAV serotype may be, or have, a
mutation in the AAV9 sequence as described by N Pulicherla et al.
(Molecular Therapy 19(6): 1070-1078 (2011), herein incorporated by
reference in its entirety), such as but not limited to, AAV9.9,
AAV9.11, AAV9.13, AAV9.16, AAV9.24, AAV9.45, AAV9.47, AAV9.61,
AAV9.68, AAV9.84.
[0038] In some embodiments, the AAV serotype may be, or have, a
sequence as described in U.S. Pat. No. 6,156,303, the contents of
which are herein incorporated by reference in their entirety, such
as, but not limited to, AAV3B (SEQ ID NO: 1 and 10 of U.S. Pat. No.
6,156,303), AAV6 (SEQ ID NO: 2, 7 and 11 of U.S. Pat. No.
6,156,303), AAV2 (SEQ ID NO: 3 and 8 of U.S. Pat. No. 6,156,303),
AAV3A (SEQ ID NO: 4 and 9, of U.S. Pat. No. 6,156,303), or
derivatives thereof.
[0039] In some embodiments, the AAV serotype may be, or have, a
sequence as described in United States Publication No.
US20140359799, the contents of which are herein incorporated by
reference in their entirety, such as, but not limited to, AAV8 (SEQ
ID NO: 1 of US20140359799), AAVDJ (SEQ ID NO: 2 and 3 of
US20140359799), or variants thereof.
[0040] In some embodiments, the serotype may be AAVDJ or a variant
thereof, such as AAVDJ8 (or AAV-DJ8), as described by Grimm et al.
(Journal of Virology 82(12): 5887-5911 (2008), herein incorporated
by reference in its entirety). The amino acid sequence of AAVDJ8
may comprise two or more mutations in order to remove the heparin
binding domain (HBD). As a non-limiting example, the AAV-DJ
sequence described as SEQ ID NO: 1 in U.S. Pat. No. 7,588,772, the
contents of which are herein incorporated by reference in their
entirety, may comprise two mutations: (1) R587Q where arginine (R;
Arg) at amino acid 587 is changed to glutamine (Q; Gln) and (2)
R590T where arginine (R; Arg) at amino acid 590 is changed to
threonine (T; Thr). As another non-limiting example, may comprise
three mutations: (1) K406R where lysine (K; Lys) at amino acid 406
is changed to arginine (R; Arg), (2) R587Q where arginine (R; Arg)
at amino acid 587 is changed to glutamine (Q; Gln) and (3) R590T
where arginine (R; Arg) at amino acid 590 is changed to threonine
(T; Thr).
[0041] In some embodiments, the AAV serotype may be, or have, a
sequence of AAV4 as described in International Publication No.
WO1998011244, the contents of which are herein incorporated by
reference in their entirety, such as, but not limited to AAV4 (SEQ
ID NO: 1-20 of WO1998011244).
[0042] In some embodiments, the AAV serotype may be, or have, a
mutation in the AAV2 sequence to generate AAV2G9 as described in
International Publication No. WO2014144229 and herein incorporated
by reference in its entirety.
[0043] In some embodiments, the AAV serotype may be, or have, a
sequence as described in International Publication No.
WO2005033321, the contents of which are herein incorporated by
reference in their entirety, such as, but not limited to AAV3-3
(SEQ ID NO: 217 of WO2005033321), AAV1 (SEQ ID NO: 219 and 202 of
WO2005033321), AAV106.1/hu.37 (SEQ ID No: 10 of WO2005033321),
AAV114.3/hu.40 (SEQ ID No: 11 of WO2005033321), AAV127.2/hu.41 (SEQ
ID NO:6 and 8 of WO2005033321), AAV128.3/hu.44 (SEQ ID No: 81 of
WO2005033321), AAV130.4/hu.48 (SEQ ID NO: 78 of WO2005033321),
AAV145.1/hu.53 (SEQ ID No: 176 and 177 of WO2005033321),
AAV145.6/hu.56 (SEQ ID NO: 168 and 192 of WO2005033321),
AAV16.12/hu.11 (SEQ ID NO: 153 and 57 of WO2005033321),
AAV16.8/hu.10 (SEQ ID NO: 156 and 56 of WO2005033321),
AAV161.10/hu.60 (SEQ ID No: 170 of WO2005033321), AAV161.6/hu.61
(SEQ ID No: 174 of WO2005033321), AAV1-7/rh.48 (SEQ ID NO: 32 of
WO2005033321), AAV1-8/rh.49 (SEQ ID NOs: 103 and 25 of
WO2005033321), AAV2 (SEQ ID NO: 211 and 221 of WO2005033321),
AAV2-15/rh.62 (SEQ ID No: 33 and 114 of WO2005033321), AAV2-3/rh.61
(SEQ ID NO: 21 of WO2005033321), AAV2-4/rh.50 (SEQ ID No: 23 and
108 of WO2005033321), AAV2-5/rh.51 (SEQ ID NO: 104 and 22 of
WO2005033321), AAV3.1/hu.6 (SEQ ID NO: 5 and 84 of WO2005033321),
AAV3.1/hu.9 (SEQ ID NO: 155 and 58 of WO2005033321), AAV3-11/rh.53
(SEQ ID NO: 186 and 176 of WO2005033321), AAV3-3 (SEQ ID NO: 200 of
WO2005033321), AAV33.12/hu.17 (SEQ ID NO:4 of WO2005033321),
AAV33.4/hu.15 (SEQ ID No: 50 of WO2005033321), AAV33.8/hu.16 (SEQ
ID No: 51 of WO2005033321), AAV3-9/rh.52 (SEQ ID NO: 96 and 18 of
WO2005033321), AAV4-19/rh.55 (SEQ ID NO: 117 of WO2005033321),
AAV4-4 (SEQ ID NO: 201 and 218 of WO2005033321), AAV4-9/rh.54 (SEQ
ID NO: 116 of WO2005033321), AAV5 (SEQ ID NO: 199 and 216 of
WO2005033321), AAV52.1/hu.20 (SEQ ID NO: 63 of WO2005033321),
AAV52/hu.19 (SEQ ID NO: 133 of WO2005033321), AAV5-22/rh.58 (SEQ ID
No: 27 of WO2005033321), AAV5-3/rh.57 (SEQ ID NO: 105 of
WO2005033321), AAV5-3/rh.57 (SEQ ID No: 26 of WO2005033321),
AAV58.2/hu.25 (SEQ ID No: 49 of WO2005033321), AAV6 (SEQ ID NO: 203
and 220 of WO2005033321), AAV7 (SEQ ID NO: 222 and 213 of
WO2005033321), AAV7.3/hu.7 (SEQ ID No: 55 of WO2005033321), AAV8
(SEQ ID NO: 223 and 214 of WO2005033321), AAVH-1/hu.1 (SEQ ID No:
46 of WO2005033321), AAVH-5/hu.3 (SEQ ID No: 44 of WO2005033321),
AAVhu.1 (SEQ ID NO: 144 of WO2005033321), AAVhu.10 (SEQ ID NO: 156
of WO2005033321), AAVhu.11 (SEQ ID NO: 153 of WO2005033321),
AAVhu.12 (WO2005033321 SEQ ID NO: 59), AAVhu. 13 (SEQ ID NO: 129 of
WO2005033321), AAVhu.14/AAV9 (SEQ ID NO: 123 and 3 of
WO2005033321), AAVhu.15 (SEQ ID NO: 147 of WO2005033321), AAVhu.16
(SEQ ID NO: 148 of WO2005033321), AAVhu.17 (SEQ ID NO: 83 of
WO2005033321), AAVhu.18 (SEQ ID NO: 149 of WO2005033321), AAVhu.19
(SEQ ID NO: 133 of WO2005033321), AAVhu.2 (SEQ ID NO: 143 of
WO2005033321), AAVhu.20 (SEQ ID NO: 134 of WO2005033321), AAVhu.21
(SEQ ID NO: 135 of WO2005033321), AAVhu.22 (SEQ ID NO: 138 of
WO2005033321), AAVhu.23.2 (SEQ ID NO: 137 of WO2005033321),
AAVhu.24 (SEQ ID NO: 136 of WO2005033321), AAVhu.25 (SEQ ID NO: 146
of WO2005033321), AAVhu.27 (SEQ ID NO: 140 of WO2005033321),
AAVhu.29 (SEQ ID NO: 132 of WO2005033321), AAVhu.3 (SEQ ID NO: 145
of WO2005033321), AAVhu.31 (SEQ ID NO: 121 of WO2005033321),
AAVhu.32 (SEQ ID NO: 122 of WO2005033321), AAVhu.34 (SEQ ID NO: 125
of WO2005033321), AAVhu.35 (SEQ ID NO: 164 of WO2005033321),
AAVhu.37 (SEQ ID NO: 88 of WO2005033321), AAVhu.39 (SEQ ID NO: 102
of WO2005033321), AAVhu.4 (SEQ ID NO: 141 of WO2005033321),
AAVhu.40 (SEQ ID NO: 87 of WO2005033321), AAVhu.41 (SEQ ID NO: 91
of WO2005033321), AAVhu.42 (SEQ ID NO: 85 of WO2005033321),
AAVhu.43 (SEQ ID NO: 160 of WO2005033321), AAVhu.44 (SEQ ID NO: 144
of WO2005033321), AAVhu.45 (SEQ ID NO: 127 of WO2005033321),
AAVhu.46 (SEQ ID NO: 159 of WO2005033321), AAVhu.47 (SEQ ID NO: 128
of WO2005033321), AAVhu.48 (SEQ ID NO: 157 of WO2005033321),
AAVhu.49 (SEQ ID NO: 189 of WO2005033321), AAVhu.51 (SEQ ID NO: 190
of WO2005033321), AAVhu.52 (SEQ ID NO: 191 of WO2005033321),
AAVhu.53 (SEQ ID NO: 186 of WO2005033321), AAVhu.54 (SEQ ID NO: 188
of WO2005033321), AAVhu.55 (SEQ ID NO: 187 of WO2005033321),
AAVhu.56 (SEQ ID NO: 192 of WO2005033321), AAVhu.57 (SEQ ID NO: 193
of WO2005033321), AAVhu.58 (SEQ ID NO: 194 of WO2005033321),
AAVhu.6 (SEQ ID NO: 84 of WO2005033321), AAVhu.60 (SEQ ID NO: 184
of WO2005033321), AAVhu.61 (SEQ ID NO: 185 of WO2005033321),
AAVhu.63 (SEQ ID NO: 195 of WO2005033321), AAVhu.64 (SEQ ID NO: 196
of WO2005033321), AAVhu.66 (SEQ ID NO: 197 of WO2005033321),
AAVhu.67 (SEQ ID NO: 198 of WO2005033321), AAVhu.7 (SEQ ID NO: 150
of WO2005033321), AAVhu.8 (WO2005033321 SEQ ID NO: 12), AAVhu.9
(SEQ ID NO: 155 of WO2005033321), AAVLG-10/rh.40 (SEQ ID No: 14 of
WO2005033321), AAVLG-4/rh.38 (SEQ ID NO: 86 of WO2005033321),
AAVLG-4/rh.38 (SEQ ID No: 7 of WO2005033321), AAVN721-8/rh.43 (SEQ
ID NO: 163 of WO2005033321), AAVN721-8/rh.43 (SEQ ID No: 43 of
WO2005033321), AAVpi.1 (WO2005033321 SEQ ID NO: 28), AAVpi.2
(WO2005033321 SEQ ID NO: 30), AAVpi.3 (WO2005033321 SEQ ID NO: 29),
AAVrh.38 (SEQ ID NO: 86 of WO2005033321), AAVrh.40 (SEQ ID NO: 92
of WO2005033321), AAVrh.43 (SEQ ID NO: 163 of WO2005033321),
AAVrh.44 (WO2005033321 SEQ ID NO: 34), AAVrh.45 (WO2005033321 SEQ
ID NO: 41), AAVrh.47 (WO2005033321 SEQ ID NO: 38), AAVrh.48 (SEQ ID
NO: 115 of WO2005033321), AAVrh.49 (SEQ ID NO: 103 of
WO2005033321), AAVrh.50 (SEQ ID NO: 108 of WO2005033321), AAVrh.51
(SEQ ID NO: 104 of WO2005033321), AAVrh.52 (SEQ ID NO: 96 of
WO2005033321), AAVrh.53 (SEQ ID NO: 97 of WO2005033321), AAVrh.55
(WO2005033321 SEQ ID NO: 37), AAVrh.56 (SEQ ID NO: 152 of
WO2005033321), AAVrh.57 (SEQ ID NO: 105 of WO2005033321), AAVrh.58
(SEQ ID NO: 106 of WO2005033321), AAVrh.59 (WO2005033321 SEQ ID NO:
42), AAVrh.60 (WO2005033321 SEQ ID NO: 31), AAVrh.61 (SEQ ID NO:
107 of WO2005033321), AAVrh.62 (SEQ ID NO: 114 of WO2005033321),
AAVrh.64 (SEQ ID NO: 99 of WO2005033321), AAVrh.65 (WO2005033321
SEQ ID NO: 35), AAVrh.68 (WO2005033321 SEQ ID NO: 16), AAVrh.69
(WO2005033321 SEQ ID NO: 39), AAVrh.70 (WO2005033321 SEQ ID NO:
20), AAVrh.72 (WO2005033321 SEQ ID NO: 9), or variants thereof
including, but not limited to, AAVcy.2, AAVcy.3, AAVcy.4, AAVcy.5,
AAVcy.6, AAVrh.12, AAVrh.17, AAVrh.18, AAVrh.19, AAVrh.21,
AAVrh.22, AAVrh.23, AAVrh.24, AAVrh.25, AAVrh.25/42 15, AAVrh.31,
AAVrh.32, AAVrh.33, AAVrh.34, AAVrh.35, AAVrh.36, AAVrh.37,
AAVrh14. Non limiting examples of variants include SEQ ID NO: 13,
15, 17, 19, 24, 36, 40, 45, 47, 48, 51-54, 60-62, 64-77, 79, 80,
82, 89, 90, 93-95, 98, 100, 101, 109-113, 118-120, 124, 126, 131,
139, 142, 151, 154, 158, 161, 162, 165-183, 202, 204-212, 215, 219,
224-236, of WO2005033321, the contents of which are herein
incorporated by reference in their entirety.
[0044] In some embodiments, the AAV serotype may be, or have, a
sequence as described in International Publication No.
WO2015168666, the contents of which are herein incorporated by
reference in their entirety, such as, but not limited to, AAVrh8R
(SEQ ID NO: 9 of WO2015168666), AAVrh8R A586R mutant (SEQ ID NO: 10
of WO2015168666), AAVrh8R R533A mutant (SEQ ID NO: 11 of
WO2015168666), or variants thereof.
[0045] In some embodiments, the AAV serotype may be, or have, a
sequence as described in U.S. Pat. No. 9,233,131, the contents of
which are herein incorporated by reference in their entirety, such
as, but not limited to, AAVhE1.1 (SEQ ID NO:44 of U.S. Pat. No.
9,233,131), AAVhEr1.5 (SEQ ID NO:45 of U.S. Pat. No. 9,233,131),
AAVhER1.14 (SEQ ID NO:46 of U.S. Pat. No. 9,233,131), AAVhEr1.8
(SEQ ID NO:47 of U.S. Pat. No. 9,233,131), AAVhEr1.16 (SEQ ID NO:48
of U.S. Pat. No. 9,233,131), AAVhEr1.18 (SEQ ID NO:49 of U.S. Pat.
No. 9,233,131), AAVhEr1.35 (SEQ ID NO:50 of U.S. Pat. No.
9,233,131), AAVhEr1.7 (SEQ ID NO:51 of U.S. Pat. No. 9,233,131),
AAVhEr1.36 (SEQ ID NO:52 of U.S. Pat. No. 9,233,131), AAVhEr2.29
(SEQ ID NO:53 of U.S. Pat. No. 9,233,131), AAVhEr2.4 (SEQ ID NO:54
of US9233131), AAVhEr2.16 (SEQ ID NO:55 of U.S. Pat. No.
9,233,131), AAVhEr2.30 (SEQ ID NO:56 of U.S. Pat. No. 9,233,131),
AAVhEr2.31 (SEQ ID NO:58 of U.S. Pat. No. 9,233,131), AAVhEr2.36
(SEQ ID NO:57 of U.S. Pat. No. 9,233,131), AAVhER1.23 (SEQ ID NO:53
of U.S. Pat. No. 9,233,131), AAVhEr3.1 (SEQ ID NO:59 of US9233131),
AAV2.5T (SEQ ID NO:42 of U.S. Pat. No. 9,233,131), or variants
thereof.
[0046] In some embodiments, the AAV serotype may be, or have, a
sequence as described in United States Patent Publication No.
US20150376607, the contents of which are herein incorporated by
reference in their entirety, such as, but not limited to, AAV-PAEC
(SEQ ID NO:1 of US20150376607), AAV-LK01 (SEQ ID NO:2 of
US20150376607), AAV-LK02 (SEQ ID NO:3 of US20150376607), AAV-LK03
(SEQ ID NO:4 of US20150376607), AAV-LK04 (SEQ ID NO:5 of
US20150376607), AAV-LK05 (SEQ ID NO:6 of US20150376607), AAV-LK06
(SEQ ID NO:7 of US20150376607), AAV-LK07 (SEQ ID NO:8 of
US20150376607), AAV-LK08 (SEQ ID NO:9 of US20150376607), AAV-LK09
(SEQ ID NO:10 of US20150376607), AAV-LK 10 (SEQ ID NO:11 of
US20150376607), AAV-LK 11 (SEQ ID NO:12 of US20150376607), AAV-LK
12 (SEQ ID NO: 13 of US20150376607), AAV-LK13 (SEQ ID NO:14 of
US20150376607), AAV-LK14 (SEQ ID NO:15 of US20150376607), AAV-LK15
(SEQ ID NO:16 of US20150376607), AAV-LK16 (SEQ ID NO:17 of
US20150376607), AAV-LK17 (SEQ ID NO:18 of US20150376607), AAV-LK18
(SEQ ID NO: 19 of US20150376607), AAV-LK19 (SEQ ID NO:20 of
US20150376607), AAV-PAEC2 (SEQ ID NO:21 of US20150376607),
AAV-PAEC4 (SEQ ID NO:22 of US20150376607), AAV-PAEC6 (SEQ ID NO:23
of US20150376607), AAV-PAEC7 (SEQ ID NO:24 of US20150376607),
AAV-PAEC8 (SEQ ID NO:25 of US20150376607), AAV-PAEC11 (SEQ ID NO:26
of US20150376607), AAV-PAEC12 (SEQ ID NO:27, of US20150376607), or
variants thereof.
[0047] In some embodiments, the AAV serotype may be, or have, a
sequence as described in U.S. Pat. No. 9,163,261, the contents of
which are herein incorporated by reference in their entirety, such
as, but not limited to, AAV-2-pre-miRNA-101 (SEQ ID NO: 1 U.S. Pat.
No. 9,163,261), or variants thereof.
[0048] In some embodiments, the AAV serotype may be, or have, a
sequence as described in United States Patent Publication No.
US20150376240, the contents of which are herein incorporated by
reference in their entirety, such as, but not limited to, AAV-8h
(SEQ ID NO: 6 of US20150376240), AAV-8b (SEQ ID NO: 5 of
US20150376240), AAV-h (SEQ ID NO: 2 of US20150376240), AAV-b (SEQ
ID NO: 1 of US20150376240), or variants thereof.
[0049] In some embodiments, the AAV serotype may be, or have, a
sequence as described in United States Patent Publication No.
US20160017295, the contents of which are herein incorporated by
reference in their entirety, such as, but not limited to, AAV SM
10-2 (SEQ ID NO: 22 of US20160017295), AAV Shuffle 100-1 (SEQ ID
NO: 23 of US20160017295), AAV Shuffle 100-3 (SEQ ID NO: 24 of
US20160017295), AAV Shuffle 100-7 (SEQ ID NO: 25 of US20160017295),
AAV Shuffle 10-2 (SEQ ID NO: 34 of US20160017295), AAV Shuffle 10-6
(SEQ ID NO: 35 of US20160017295), AAV Shuffle 10-8 (SEQ ID NO: 36
of US20160017295), AAV Shuffle 100-2 (SEQ ID NO: 37 of
US20160017295), AAV SM 10-1 (SEQ ID NO: 38 of US20160017295), AAV
SM 10-8 (SEQ ID NO: 39 of US20160017295), AAV SM 100-3 (SEQ ID NO:
40 of US20160017295), AAV SM 100-10 (SEQ ID NO: 41 of
US20160017295), or variants thereof.
[0050] In some embodiments, the AAV serotype may be, or have, a
sequence as described in United States Patent Publication No.
US20150238550, the contents of which are herein incorporated by
reference in their entirety, such as, but not limited to, BNP61 AAV
(SEQ ID NO: 1 of US20150238550), BNP62 AAV (SEQ ID NO: 3 of
US20150238550), BNP63 AAV (SEQ ID NO: 4 of US20150238550), or
variants thereof.
[0051] In some embodiments, the AAV serotype may be or may have a
sequence as described in United States Patent Publication No.
US20150315612, the contents of which are herein incorporated by
reference in their entirety, such as, but not limited to, AAVrh.50
(SEQ ID NO: 108 of US20150315612), AAVrh.43 (SEQ ID NO: 163 of
US20150315612), AAVrh.62 (SEQ ID NO: 114 of US20150315612),
AAVrh.48 (SEQ ID NO: 115 of US20150315612), AAVhu.19 (SEQ ID NO:
133 of US20150315612), AAVhu.11 (SEQ ID NO: 153 of US20150315612),
AAVhu.53 (SEQ ID NO: 186 of US20150315612), AAV4-8/rh.64 (SEQ ID
No: 15 of US20150315612), AAVLG-9/hu.39 (SEQ ID No: 24 of
US20150315612), AAV54.5/hu.23 (SEQ ID No: 60 of US20150315612),
AAV54.2/hu.22 (SEQ ID No: 67 of US20150315612), AAV54.7/hu.24 (SEQ
ID No: 66 of US20150315612), AAV54.1/hu.21 (SEQ ID No: 65 of
US20150315612), AAV54.4R/hu.27 (SEQ ID No: 64 of US20150315612),
AAV46.2/hu.28 (SEQ ID No: 68 of US20150315612), AAV46.6/hu.29 (SEQ
ID No: 69 of US20150315612), AAV128.1/hu.43 (SEQ ID No: 80 of
US20150315612), or variants thereof.
[0052] In some embodiments, the AAV serotype may be, or have, a
sequence as described in International Publication No.
WO2015121501, the contents of which are herein incorporated by
reference in their entirety, such as, but not limited to, true type
AAV (ttAAV) (SEQ ID NO: 2 of WO2015121501), "UPenn AAV10" (SEQ ID
NO: 8 of WO2015121501), "Japanese AAV10" (SEQ ID NO: 9 of
WO2015121501), or variants thereof.
[0053] According to the present disclosure, AAV capsid serotype
selection or use may be from a variety of species. In one
embodiment, the AAV may be an avian AAV (AAAV). The AAAV serotype
may be, or have, a sequence as described in U.S. Pat. No.
9,238,800, the contents of which are herein incorporated by
reference in their entirety, such as, but not limited to, AAAV (SEQ
ID NO: 1, 2, 4, 6, 8, 10, 12, and 14 of U.S. Pat. No. 9,238,800),
or variants thereof.
[0054] In one embodiment, the AAV may be a bovine AAV (BAAV). The
BAAV serotype may be, or have, a sequence as described in U.S. Pat.
No. 9,193,769, the contents of which are herein incorporated by
reference in their entirety, such as, but not limited to, BAAV (SEQ
ID NO: 1 and 6 of U.S. Pat. No. 9,193,769), or variants thereof.
The BAAV serotype may be or have a sequence as described in U.S.
Pat. No. 7,427,396, the contents of which are herein incorporated
by reference in their entirety, such as, but not limited to, BAAV
(SEQ ID NO: 5 and 6 of U.S. Pat. No. 7,427,396), or variants
thereof.
[0055] In one embodiment, the AAV may be a caprine AAV. The caprine
AAV serotype may be, or have, a sequence as described in U.S. Pat.
No. 7,427,396, the contents of which are herein incorporated by
reference in their entirety, such as, but not limited to, caprine
AAV (SEQ ID NO: 3 of U.S. Pat. No. 7,427,396), or variants
thereof.
[0056] In other embodiments the AAV may be engineered as a hybrid
AAV from two or more parental serotypes. In one embodiment, the AAV
may be AAV2G9 which comprises sequences from AAV2 and AAV9. The
AAV2G9 AAV serotype may be, or have, a sequence as described in
United States Patent Publication No. US20160017005, the contents of
which are herein incorporated by reference in its entirety.
[0057] In one embodiment, the AAV may be a serotype generated by
the AAV9 capsid library with mutations in amino acids 390-627 (VP1
numbering) as described by Pulicherla et al. (Molecular Therapy
19(6): 1070-1078 (2011), the contents of which are herein
incorporated by reference in their entirety. The serotype and
corresponding nucleotide and amino acid substitutions may be, but
is not limited to, AAV9.1 (G1594C; D532H), AAV6.2 (T1418A and
T1436X; V473D and I479K), AAV9.3 (T1238A; F413Y), AAV9.4 (T1250C
and A1617T; F417S), AAV9.5 (A1235G, A1314T, A1642G, C1760T; Q412R,
T548A, A587V), AAV9.6 (T1231A; F411I), AAV9.9 (G1203A, G1785T;
W595C), AAV9.10 (A1500G, T1676C; M559T), AAV9.11 (A1425T, A1702C,
A1769T; T568P, Q590L), AAV9.13 (A1369C, A1720T; N457H, T574S),
AAV9.14 (T1340A, T1362C, T1560C, G1713A; L447H), AAV9.16 (A1775T;
Q592L), AAV9.24 (T1507C, T1521G; W503R), AAV9.26 (A1337G, A1769C;
Y446C, Q590P), AAV9.33 (A1667C; D556A), AAV9.34 (A1534G, C1794T;
N512D), AAV9.35 (A1289T, T1450A, C1494T, A1515T, C1794A, G1816A;
Q430L, Y484N, N98K, V606I), AAV9.40 (A1694T, E565V), AAV9.41
(A1348T, T1362C; T450S), AAV9.44 (A1684C, A1701T, A1737G; N562H,
K567N), AAV9.45 (A1492T, C1804T; N498Y, L602F), AAV9.46 (G1441C,
T1525C, T1549G; G481R, W509R, L517V), 9.47 (G1241A, G1358A, A1669G,
C1745T; S414N, G453D, K557E, T582I), AAV9.48 (C1445T, A1736T;
P482L, Q579L), AAV9.50 (A1638T, C1683T, T1805A; Q546H, L602H),
AAV9.53 (G1301A, A1405C, C1664T, G1811T; R134Q, S469R, A555V,
G604V), AAV9.54 (C1531A, T1609A; L511I, L537M), AAV9.55 (T1605A;
F535L), AAV9.58 (C1475T, C1579A; T492I, H527N), AAV.59 (T1336C;
Y446H), AAV9.61 (A1493T; N498I), AAV9.64 (C1531A, A1617T; L511I),
AAV9.65 (C1335T, T1530C, C1568A; A523D), AAV9.68 (C1510A; P504T),
AAV9.80 (G1441A; G481R), AAV9.83 (C1402A, A1500T; P468T, E500D),
AAV9.87 (T1464C, T1468C; S490P), AAV9.90 (A1196T; Y399F), AAV9.91
(T1316G, A1583T, C1782G, T1806C; L439R, K528I), AAV9.93 (A1273G,
A1421G, A1638C, C1712T, G1732A, A1744T, A1832T; S425G, Q474R,
Q546H, P571L, G578R, T582S, D611V), AAV9.94 (A1675T; M559L) and
AAV9.95 (T1605A; F535L).
[0058] In some embodiments, the AAV serotype may be, or have, a
sequence as described in International Publication No.
WO2016049230, the contents of which are herein incorporated by
reference in their entirety, such as, but not limited to AAVF
1/HSC1 (SEQ ID NO: 2 and 20 of WO2016049230), AAVF2/HSC2 (SEQ ID
NO: 3 and 21 of WO2016049230), AAVF3/HSC3 (SEQ ID NO: 5 and 22 of
WO2016049230), AAVF4/HSC4 (SEQ ID NO: 6 and 23 of WO2016049230),
AAVF5/HSC5 (SEQ ID NO: 11 and 25 of WO2016049230), AAVF6/HSC6 (SEQ
ID NO: 7 and 24 of WO2016049230), AAVF7/HSC7 (SEQ ID NO: 8 and 27
of WO2016049230), AAVF8/HSC8 (SEQ ID NO: 9 and 28 of WO2016049230),
AAVF9/HSC9 (SEQ ID NO: 10 and 29 of WO2016049230), AAVF11/HSC11
(SEQ ID NO: 4 and 26 of WO2016049230), AAVF12/HSC12 (SEQ ID NO: 12
and 30 of WO2016049230), AAVF13/HSC13 (SEQ ID NO: 14 and 31 of
WO2016049230), AAVF14/HSC14 (SEQ ID NO: 15 and 32 of WO2016049230),
AAVF15/HSC15 (SEQ ID NO: 16 and 33 of WO2016049230), AAVF16/HSC16
(SEQ ID NO: 17 and 34 of WO2016049230), AAVF17/HSC17 (SEQ ID NO: 13
and 35 of WO2016049230), or variants or derivatives thereof.
[0059] In some embodiments, the AAV serotype may be, or have, a
sequence as described in U.S. Pat. No. 8,734,809, the contents of
which are herein incorporated by reference in their entirety, such
as, but not limited to, AAV CBr-E1 (SEQ ID NO: 13 and 87 of U.S.
Pat. No. 8,734,809), AAV CBr-E2 (SEQ ID NO: 14 and 88 of U.S. Pat.
No. 8,734,809), AAV CBr-E3 (SEQ ID NO: 15 and 89 of U.S. Pat. No.
8,734,809), AAV CBr-E4 (SEQ ID NO: 16 and 90 of U.S. Pat. No.
8,734,809), AAV CBr-E5 (SEQ ID NO: 17 and 91 of U.S. Pat. No.
8,734,809), AAV CBr-e5 (SEQ ID NO: 18 and 92 of U.S. Pat. No.
8,734,809), AAV CBr-E6 (SEQ ID NO: 19 and 93 of U.S. Pat. No.
8,734,809), AAV CBr-E7 (SEQ ID NO: 20 and 94 of U.S. Pat. No.
8,734,809), AAV CBr-E8 (SEQ ID NO: 21 and 95 of U.S. Pat. No.
8,734,809), AAV CLv-D1 (SEQ ID NO: 22 and 96 of U.S. Pat. No.
8,734,809), AAV CLv-D2 (SEQ ID NO: 23 and 97 of U.S. Pat. No.
8,734,809), AAV CLv-D3 (SEQ ID NO: 24 and 98 of U.S. Pat. No.
8,734,809), AAV CLv-D4 (SEQ ID NO: 25 and 99 of U.S. Pat. No.
8,734,809), AAV CLv-D5 (SEQ ID NO: 26 and 100 of U.S. Pat. No.
8,734,809), AAV CLv-D6 (SEQ ID NO: 27 and 101 of U.S. Pat. No.
8,734,809), AAV CLv-D7 (SEQ ID NO: 28 and 102 of U.S. Pat. No.
8,734,809), AAV CLv-D8 (SEQ ID NO: 29 and 103 of U.S. Pat. No.
8,734,809), AAV CLv-E1 (SEQ ID NO: 13 and 87 of U.S. Pat. No.
8,734,809), AAV CLv-R1 (SEQ ID NO: 30 and 104 of U.S. Pat. No.
8,734,809), AAV CLv-R2 (SEQ ID NO: 31 and 105 of U.S. Pat. No.
8,734,809), AAV CLv-R3 (SEQ ID NO: 32 and 106 of U.S. Pat. No.
8,734,809), AAV CLv-R4 (SEQ ID NO: 33 and 107 of U.S. Pat. No.
8,734,809), AAV CLv-R5 (SEQ ID NO: 34 and 108 of U.S. Pat. No.
8,734,809), AAV CLv-R6 (SEQ ID NO: 35 and 109 of U.S. Pat. No.
8,734,809), AAV CLv-R7 (SEQ ID NO: 36 and 110 of U.S. Pat. No.
8,734,809), AAV CLv-R8 (SEQ ID NO: X and X of U.S. Pat. No.
8,734,809), AAV CLv-R9 (SEQ ID NO: X and X of U.S. Pat. No.
8,734,809), AAV CLg-F1 (SEQ ID NO: 39 and 113 of U.S. Pat. No.
8,734,809), AAV CLg-F2 (SEQ ID NO: 40 and 114 of U.S. Pat. No.
8,734,809), AAV CLg-F3 (SEQ ID NO: 41 and 115 of U.S. Pat. No.
8,734,809), AAV CLg-F4 (SEQ ID NO: 42 and 116 of U.S. Pat. No.
8,734,809), AAV CLg-F5 (SEQ ID NO: 43 and 117 of U.S. Pat. No.
8,734,809), AAV CLg-F6 (SEQ ID NO: 43 and 117 of U.S. Pat. No.
8,734,809), AAV CLg-F7 (SEQ ID NO: 44 and 118 of U.S. Pat. No.
8,734,809), AAV CLg-F8 (SEQ ID NO: 43 and 117 of U.S. Pat. No.
8,734,809), AAV CSp-1 (SEQ ID NO: 45 and 119 of U.S. Pat. No.
8,734,809), AAV CSp-10 (SEQ ID NO: 46 and 120 of U.S. Pat. No.
8,734,809), AAV CSp-11 (SEQ ID NO: 47 and 121 of U.S. Pat. No.
8,734,809), AAV CSp-2 (SEQ ID NO: 48 and 122 of U.S. Pat. No.
8,734,809), AAV CSp-3 (SEQ ID NO: 49 and 123 of U.S. Pat. No.
8,734,809), AAV CSp-4 (SEQ ID NO: 50 and 124 of U.S. Pat. No.
8,734,809), AAV CSp-6 (SEQ ID NO: 51 and 125 of U.S. Pat. No.
8,734,809), AAV CSp-7 (SEQ ID NO: 52 and 126 of U.S. Pat. No.
8,734,809), AAV CSp-8 (SEQ ID NO: 53 and 127 of U.S. Pat. No.
8,734,809), AAV CSp-9 (SEQ ID NO: 54 and 128 of U.S. Pat. No.
8,734,809), AAV CHt-2 (SEQ ID NO: 55 and 129 of U.S. Pat. No.
8,734,809), AAV CHt-3 (SEQ ID NO: 56 and 130 of U.S. Pat. No.
8,734,809), AAV CKd-1 (SEQ ID NO: 57 and 131 of U.S. Pat. No.
8,734,809), AAV CKd-10 (SEQ ID NO: 58 and 132 of U.S. Pat. No.
8,734,809), AAV CKd-2 (SEQ ID NO: 59 and 133 of U.S. Pat. No.
8,734,809), AAV CKd-3 (SEQ ID NO: 60 and 134 of U.S. Pat. No.
8,734,809), AAV CKd-4 (SEQ ID NO: 61 and 135 of U.S. Pat. No.
8,734,809), AAV CKd-6 (SEQ ID NO: 62 and 136 of U.S. Pat. No.
8,734,809), AAV CKd-7 (SEQ ID NO: 63 and 137 of U.S. Pat. No.
8,734,809), AAV CKd-8 (SEQ ID NO: 64 and 138 of U.S. Pat. No.
8,734,809), AAV CLv-1 (SEQ ID NO: 35 and 139 of U.S. Pat. No.
8,734,809), AAV CLv-12 (SEQ ID NO: 66 and 140 of U.S. Pat. No.
8,734,809), AAV CLv-13 (SEQ ID NO: 67 and 141 of U.S. Pat. No.
8,734,809), AAV CLv-2 (SEQ ID NO: 68 and 142 of U.S. Pat. No.
8,734,809), AAV CLv-3 (SEQ ID NO: 69 and 143 of U.S. Pat. No.
8,734,809), AAV CLv-4 (SEQ ID NO: 70 and 144 of U.S. Pat. No.
8,734,809), AAV CLv-6 (SEQ ID NO: 71 and 145 of U.S. Pat. No.
8,734,809), AAV CLv-8 (SEQ ID NO: 72 and 146 of U.S. Pat. No.
8,734,809), AAV CKd-B1 (SEQ ID NO: 73 and 147 of U.S. Pat. No.
8,734,809), AAV CKd-B2 (SEQ ID NO: 74 and 148 of U.S. Pat. No.
8,734,809), AAV CKd-B3 (SEQ ID NO: 75 and 149 of U.S. Pat. No.
8,734,809), AAV CKd-B4 (SEQ ID NO: 76 and 150 of U.S. Pat. No.
8,734,809), AAV CKd-B5 (SEQ ID NO: 77 and 151 of U.S. Pat. No.
8,734,809), AAV CKd-B6 (SEQ ID NO: 78 and 152 of U.S. Pat. No.
8,734,809), AAV CKd-B7 (SEQ ID NO: 79 and 153 of U.S. Pat. No.
8,734,809), AAV CKd-B8 (SEQ ID NO: 80 and 154 of U.S. Pat. No.
8,734,809), AAV CKd-H1 (SEQ ID NO: 81 and 155 of U.S. Pat. No.
8,734,809), AAV CKd-H2 (SEQ ID NO: 82 and 156 of U.S. Pat. No.
8,734,809), AAV CKd-H3 (SEQ ID NO: 83 and 157 of U.S. Pat. No.
8,734,809), AAV CKd-H4 (SEQ ID NO: 84 and 158 of U.S. Pat. No.
8,734,809), AAV CKd-H5 (SEQ ID NO: 85 and 159 of U.S. Pat. No.
8,734,809), AAV CKd-H6 (SEQ ID NO: 77 and 151 of U.S. Pat. No.
8,734,809), AAV CHt-1 (SEQ ID NO: 86 and 160 of U.S. Pat. No.
8,734,809), AAV CLv1-1 (SEQ ID NO: 171 of U.S. Pat. No. 8,734,809),
AAV CLv1-2 (SEQ ID NO: 172 of U.S. Pat. No. 8,734,809), AAV CLv1-3
(SEQ ID NO: 173 of U.S. Pat. No. 8,734,809), AAV CLv1-4 (SEQ ID NO:
174 of U.S. Pat. No. 8,734,809), AAV Clv1-7 (SEQ ID NO: 175 of U.S.
Pat. No. 8,734,809), AAV Clv1-8 (SEQ ID NO: 176 of US8734809), AAV
Clv1-9 (SEQ ID NO: 177 of US8734809), AAV Clv1-10 (SEQ ID NO: 178
of U.S. Pat. No. 8,734,809), AAV.VR-355 (SEQ ID NO: 181 of U.S.
Pat. No. 8,734,809), AAV.hu.48R3 (SEQ ID NO: 183 of U.S. Pat. No.
8,734,809), or variants or derivatives thereof.
[0060] In some embodiments, the AAV serotype may be, or have, a
sequence as described in International Publication No.
WO2016065001, the contents of which are herein incorporated by
reference in their entirety, such as, but not limited to AAV CHt-P2
(SEQ ID NO: 1 and 51 of WO2016065001), AAV CHt-P5 (SEQ ID NO: 2 and
52 of WO2016065001), AAV CHt-P9 (SEQ ID NO: 3 and 53 of
WO2016065001), AAV CBr-7.1 (SEQ ID NO: 4 and 54 of WO2016065001),
AAV CBr-7.2 (SEQ ID NO: 5 and 55 of WO2016065001), AAV CBr-7.3 (SEQ
ID NO: 6 and 56 of WO2016065001), AAV CBr-7.4 (SEQ ID NO: 7 and 57
of WO2016065001), AAV CBr-7.5 (SEQ ID NO: 8 and 58 of
WO2016065001), AAV CBr-7.7 (SEQ ID NO: 9 and 59 of WO2016065001),
AAV CBr-7.8 (SEQ ID NO: 10 and 60 of WO2016065001), AAV CBr-7.10
(SEQ ID NO: 11 and 61 of WO2016065001), AAV CKd-N3 (SEQ ID NO: 12
and 62 of WO2016065001), AAV CKd-N4 (SEQ ID NO: 13 and 63 of
WO2016065001), AAV CKd-N9 (SEQ ID NO: 14 and 64 of WO2016065001),
AAV CLv-L4 (SEQ ID NO: 15 and 65 of WO2016065001), AAV CLv-L5 (SEQ
ID NO: 16 and 66 of WO2016065001), AAV CLv-L6 (SEQ ID NO: 17 and 67
of WO2016065001), AAV CLv-K1 (SEQ ID NO: 18 and 68 of
WO2016065001), AAV CLv-K3 (SEQ ID NO: 19 and 69 of WO2016065001),
AAV CLv-K6 (SEQ ID NO: 20 and 70 of WO2016065001), AAV CLv-M1 (SEQ
ID NO: 21 and 71 of WO2016065001), AAV CLv-M11 (SEQ ID NO: 22 and
72 of WO2016065001), AAV CLv-M2 (SEQ ID NO: 23 and 73 of
WO2016065001), AAV CLv-M5 (SEQ ID NO: 24 and 74 of WO2016065001),
AAV CLv-M6 (SEQ ID NO: 25 and 75 of WO2016065001), AAV CLv-M7 (SEQ
ID NO: 26 and 76 of WO2016065001), AAV CLv-M8 (SEQ ID NO: 27 and 77
of WO2016065001), AAV CLv-M9 (SEQ ID NO: 28 and 78 of
WO2016065001), AAV CHt-P1 (SEQ ID NO: 29 and 79 of WO2016065001),
AAV CHt-P6 (SEQ ID NO: 30 and 80 of WO2016065001), AAV CHt-P8 (SEQ
ID NO: 31 and 81 of WO2016065001), AAV CHt-6.1 (SEQ ID NO: 32 and
82 of WO2016065001), AAV CHt-6.10 (SEQ ID NO: 33 and 83 of
WO2016065001), AAV CHt-6.5 (SEQ ID NO: 34 and 84 of WO2016065001),
AAV CHt-6.6 (SEQ ID NO: 35 and 85 of WO2016065001), AAV CHt-6.7
(SEQ ID NO: 36 and 86 of WO2016065001), AAV CHt-6.8 (SEQ ID NO: 37
and 87 of WO2016065001), AAV CSp-8.10 (SEQ ID NO: 38 and 88 of
WO2016065001), AAV CSp-8.2 (SEQ ID NO: 39 and 89 of WO2016065001),
AAV CSp-8.4 (SEQ ID NO: 40 and 90 of WO2016065001), AAV CSp-8.5
(SEQ ID NO: 41 and 91 of WO2016065001), AAV CSp-8.6 (SEQ ID NO: 42
and 92 of WO2016065001), AAV CSp-8.7 (SEQ ID NO: 43 and 93 of
WO2016065001), AAV CSp-8.8 (SEQ ID NO: 44 and 94 of WO2016065001),
AAV CSp-8.9 (SEQ ID NO: 45 and 95 of WO2016065001), AAV CBr-B7.3
(SEQ ID NO: 46 and 96 of WO2016065001), AAV CBr-B7.4 (SEQ ID NO: 47
and 97 of WO2016065001), AAV3B (SEQ ID NO: 48 and 98 of
WO2016065001), AAV4 (SEQ ID NO: 49 and 99 of WO2016065001), AAV5
(SEQ ID NO: 50 and 100 of WO2016065001), or variants or derivatives
thereof.
[0061] In some embodiments, the AAV serotype may be, or have, a
modification as described in United States Publication No. US
20160361439, the contents of which are herein incorporated by
reference in their entirety, such as but not limited to, Y252F,
Y272F, Y444F, Y500F, Y700F, Y704F, Y730F, Y275F, Y281F, Y508F,
Y576F, Y612G, Y673F, and Y720F of the wild-type AAV1, AAV2, AAV3,
AAV4, AAV5, AAV6, AAV7, AAV8, AAV9, AAV10, AAV11, AAV12, and
hybrids thereof.
[0062] In some embodiments, the AAV serotype may be, or have, a
mutation as described in U.S. Pat. No. 9,546,112, the contents of
which are herein incorporated by reference in their entirety, such
as, but not limited to, at least two, but not all the F129L, D418E,
K531E, L584F, V598A and H642N mutations in the sequence of AAV6
(SEQ ID NO:4 of U.S. Pat. No. 9,546,112), AAV1 (SEQ ID NO:6 of U.S.
Pat. No. 9,546,112), AAV2, AAV3, AAV4, AAV5, AAV7, AAV9, AAV10 or
AAV11 or derivatives thereof. In yet another embodiment, the AAV
serotype may be, or have, an AAV6 sequence comprising the K531E
mutation (SEQ ID NO:5 of U.S. Pat. No. 9,546,112).
[0063] In some embodiments, the AAV serotype may be, or have, a
mutation in the AAV1 sequence, as described in in United States
Publication No. US 20130224836, the contents of which are herein
incorporated by reference in their entirety, such as, but not
limited to, at least one of the surface-exposed tyrosine residues,
preferably, at positions 252, 273, 445, 701, 705 and 731 of AAV1
(SEQ ID NO: 2 of US 20130224836) substituted with another amino
acid, preferably with a phenylalanine residue. In one embodiment,
the AAV serotype may be, or have, a mutation in the AAV9 sequence,
such as, but not limited to, at least one of the surface-exposed
tyrosine residues, preferably, at positions 252, 272, 444, 500,
700, 704 and 730 of AAV2 (SEQ ID NO: 4 of US 20130224836)
substituted with another amino acid, preferably with a
phenylalanine residue. In one embodiment, the tyrosine residue at
position 446 of AAV9 (SEQ ID NO: 6 US 20130224836) is substituted
with a phenylalanine residue.
[0064] In some embodiments, the serotype may be AAV2 or a variant
thereof, as described in International Publication No.
WO2016130589, herein incorporated by reference in its entirety. The
amino acid sequence of AAV2 may comprise N587A, E548A, or N708A
mutations. In one embodiment, the amino acid sequence of any AAV
may comprise a V708K mutation.
[0065] In one embodiment, the AAV may be a serotype selected from
any of those found in Table 1.
[0066] In one embodiment, the AAV may comprise a sequence, fragment
or variant thereof, of the sequences in Table 1.
[0067] In one embodiment, the AAV may be encoded by a sequence,
fragment or variant as described in Table 1.
TABLE-US-00001 TABLE 1 AAV Serotypes SEQ ID Serotype NO Reference
Information AAVPHP.B or 1 WO2015038958 SEQ ID NO: 8 G2B-26 and 13
AAVPHP.B 2 WO2015038958 SEQ ID NO: 9 AAVG2B-13 3 WO2015038958 SEQ
ID NO: 12 AAVTH1.1-32 4 WO2015038958 SEQ ID NO: 14 AAVTH1.1-35 5
WO2015038958 SEQ ID NO: 15 AAV1 6 US20150159173 SEQ ID NO: 11,
US20150315612 SEQ ID NO: 202 AAV1 7 US20160017295 SEQ ID NO:
1US20030138772 SEQ ID NO: 64, US20150159173 SEQ ID NO: 27,
US20150315612 SEQ ID NO: 219, U.S. Pat. No. 7,198,951 SEQ ID NO: 5
AAV1 8 US20030138772 SEQ ID NO: 6 AAV1.3 9 US20030138772 SEQ ID NO:
14 AAV10 10 US20030138772 SEQ ID NO: 117 AAV10 11 WO2015121501 SEQ
ID NO: 9 AAV10 12 WO2015121501 SEQ ID NO: 8 AAV11 13 US20030138772
SEQ ID NO: 118 AAV12 14 US20030138772 SEQ ID NO: 119 AAV2 15
US20150159173 SEQ ID NO: 7, US20150315612 SEQ ID NO: 211 AAV2 16
US20030138772 SEQ ID NO: 70, US20150159173 SEQ ID NO: 23,
US20150315612 SEQ ID NO: 221, US20160017295 SEQ ID NO: 2, U.S. Pat.
No. 6,156,303 SEQ ID NO: 4, U.S. Pat. No. 7,198,951 SEQ ID NO: 4,
WO2015121501 SEQ ID NO: 1 AAV2 17 U.S. Pat. No. 6,156,303 SEQ ID
NO: 8 AAV2 18 US20030138772 SEQ ID NO: 7 AAV2 19 U.S. Pat. No.
6,156,303 SEQ ID NO: 3 AAV2.5T 20 U.S. Pat. No. 9,233,131 SEQ ID
NO: 42 AAV223.10 21 US20030138772 SEQ ID NO: 75 AAV223.2 22
US20030138772 SEQ ID NO: 49 AAV223.2 23 US20030138772 SEQ ID NO: 76
AAV223.4 24 US20030138772 SEQ ID NO: 50 AAV223.4 25 US20030138772
SEQ ID NO: 73 AAV223.5 26 US20030138772 SEQ ID NO: 51 AAV223.5 27
US20030138772 SEQ ID NO: 74 AAV223.6 28 US20030138772 SEQ ID NO: 52
AAV223.6 29 US20030138772 SEQ ID NO: 78 AAV223.7 30 US20030138772
SEQ ID NO: 53 AAV223.7 31 US20030138772 SEQ ID NO: 77 AAV29.3 32
US20030138772 SEQ ID NO: 82 AAV29.4 33 US20030138772 SEQ ID NO: 12
AAV29.5 34 US20030138772 SEQ ID NO: 83 AAV29.5 35 US20030138772 SEQ
ID NO: 13 (AAVbb.2) AAV3 36 US20150159173 SEQ ID NO: 12 AAV3 37
US20030138772 SEQ ID NO: 71, US20150159173 SEQ ID NO: 28,
US20160017295 SEQ ID NO: 3, U.S. Pat. No. 7,198,951 SEQ ID NO: 6
AAV3 38 US20030138772 SEQ ID NO: 8 AAV3.3b 39 US20030138772 SEQ ID
NO: 72 AAV3-3 40 US20150315612 SEQ ID NO: 200 AAV3-3 41
US20150315612 SEQ ID NO: 217 AAV3a 42 US6156303 SEQ ID NO: 5 AAV3a
43 US6156303 SEQ ID NO: 9 AAV3b 44 US6156303 SEQ ID NO: 6 AAV3b 45
US6156303 SEQ ID NO: 10 AAV3b 46 US6156303 SEQ ID NO: 1 AAV4 47
US20140348794 SEQ ID NO: 17 AAV4 48 US20140348794 SEQ ID NO: 5 AAV4
49 US20140348794 SEQ ID NO: 3 AAV4 50 US20140348794 SEQ ID NO: 14
AAV4 51 US20140348794 SEQ ID NO: 15 AAV4 52 US20140348794 SEQ ID
NO: 19 AAV4 53 US20140348794 SEQ ID NO: 12 AAV4 54 US20140348794
SEQ ID NO: 13 AAV4 55 US20140348794 SEQ ID NO: 7 AAV4 56
US20140348794 SEQ ID NO: 8 AAV4 57 US20140348794 SEQ ID NO: 9 AAV4
58 US20140348794 SEQ ID NO: 2 AAV4 59 US20140348794 SEQ ID NO: 10
AAV4 60 US20140348794 SEQ ID NO: 11 AAV4 61 US20140348794 SEQ ID
NO: 18 AAV4 62 US20030138772 SEQ ID NO: 63, US20160017295 SEQ ID
NO: 4, US20140348794 SEQ ID NO: 4 AAV4 63 US20140348794 SEQ ID NO:
16 AAV4 64 US20140348794 SEQ ID NO: 20 AAV4 65 US20140348794 SEQ ID
NO: 6 AAV4 66 US20140348794 SEQ ID NO: 1 AAV42.2 67 US20030138772
SEQ ID NO: 9 AAV42.2 68 US20030138772 SEQ ID NO: 102 AAV42.3b 69
US20030138772 SEQ ID NO: 36 AAV42.3B 70 US20030138772 SEQ ID NO:
107 AAV42.4 71 US20030138772 SEQ ID NO: 33 AAV42.4 72 US20030138772
SEQ ID NO: 88 AAV42.8 73 US20030138772 SEQ ID NO: 27 AAV42.8 74
US20030138772 SEQ ID NO: 85 AAV43.1 75 US20030138772 SEQ ID NO: 39
AAV43.1 76 US20030138772 SEQ ID NO: 92 AAV43.12 77 US20030138772
SEQ ID NO: 41 AAV43.12 78 US20030138772 SEQ ID NO: 93 AAV43.20 79
US20030138772 SEQ ID NO: 42 AAV43.20 80 US20030138772 SEQ ID NO: 99
AAV43.21 81 US20030138772 SEQ ID NO: 43 AAV43.21 82 US20030138772
SEQ ID NO: 96 AAV43.23 83 US20030138772 SEQ ID NO: 44 AAV43.23 84
US20030138772 SEQ ID NO: 98 AAV43.25 85 US20030138772 SEQ ID NO: 45
AAV43.25 86 US20030138772 SEQ ID NO: 97 AAV43.5 87 US20030138772
SEQ ID NO: 40 AAV43.5 88 US20030138772 SEQ ID NO: 94 AAV4-4 89
US20150315612 SEQ ID NO: 201 AAV4-4 90 US20150315612 SEQ ID NO: 218
AAV44.1 91 US20030138772 SEQ ID NO: 46 AAV44.1 92 US20030138772 SEQ
ID NO: 79 AAV44.5 93 US20030138772 SEQ ID NO: 47 AAV44.5 94
US20030138772 SEQ ID NO: 80 AAV4407 95 US20150315612 SEQ ID NO: 90
AAV5 96 U.S. Pat. No. 7,427,396 SEQ ID NO: 1 AAV5 97 US20030138772
SEQ ID NO: 114 AAV5 98 US20160017295 SEQ ID NO: 5, U.S. Pat. No.
7,427,396 SEQ ID NO: 2, US20150315612 SEQ ID NO: 216 AAV5 99
US20150315612 SEQ ID NO: 199 AAV6 100 US20150159173 SEQ ID NO: 13
AAV6 101 US20030138772 SEQ ID NO: 65, US20150159173 SEQ ID NO: 29,
US20160017295 SEQ ID NO: 6, U.S. Pat. No. 6,156,303 SEQ ID NO: 7
AAV6 102 U.S. Pat. No. 6,156,303 SEQ ID NO: 11 AAV6 103 U.S. Pat.
No. 6,156,303 SEQ ID NO: 2 AAV6 104 US20150315612 SEQ ID NO: 203
AAV6 105 US20150315612 SEQ ID NO: 220 AAV6.1 106 US20150159173
AAV6.12 107 US20150159173 AAV6.2 108 US20150159173 AAV7 109
US20150159173 SEQ ID NO: 14 AAV7 110 US20150315612 SEQ ID NO: 183
AAV7 111 US20030138772 SEQ ID NO: 2, US20150159173 SEQ ID NO: 30,
US20150315612 SEQ ID NO: 181, US20160017295 SEQ ID NO: 7 AAV7 112
US20030138772 SEQ ID NO: 3 AAV7 113 US20030138772 SEQ ID NO: 1,
US20150315612 SEQ ID NO: 180 AAV7 114 US20150315612 SEQ ID NO: 213
AAV7 115 US20150315612 SEQ ID NO: 222 AAV8 116 US20150159173 SEQ ID
NO: 15 AAV8 117 US20150376240 SEQ ID NO: 7 AAV8 118 US20030138772
SEQ ID NO: 4, US20150315612 SEQ ID NO: 182 AAV8 119 US20030138772
SEQ ID NO: 95, US20140359799 SEQ ID NO: 1, US20150159173 SEQ ID NO:
31, US20160017295 SEQ ID NO: 8, U.S. Pat. No. 7,198,951 SEQ ID NO:
7, US20150315612 SEQ ID NO: 223 AAV8 120 US20150376240 SEQ ID NO: 8
AAV8 121 US20150315612 SEQ ID NO: 214 AAV-8b 122 US20150376240 SEQ
ID NO: 5 AAV-8b 123 US20150376240 SEQ ID NO: 3 AAV-8h 124
US20150376240 SEQ ID NO: 6 AAV-8h 125 US20150376240 SEQ ID NO: 4
AAV9 126 US20030138772 SEQ ID NO: 5 AAV9 127 U.S. Pat. No.
7,198,951 SEQ ID NO: 1 AAV9 128 US20160017295 SEQ ID NO: 9 AAV9 129
US20030138772 SEQ ID NO: 100, U.S. Pat. No. 7,198,951 SEQ ID NO: 2
AAV9 130 U.S. Pat. No. 7,198,951 SEQ ID NO: 3 AAV9 131 U.S. Pat.
No. 7,906,111 (AAVhu.14) SEQ ID NO: 3; WO2015038958 SEQ ID NO: 11
AAV9 132 U.S. Pat. No. 7,906,111 (AAVhu.14) SEQ ID NO: 123;
WO2015038958 SEQ ID NO: 2 AAVA3.1 133 US20030138772 SEQ ID NO: 120
AAVA3.3 134 US20030138772 SEQ ID NO: 57 AAVA3.3 135 US20030138772
SEQ ID NO: 66 AAVA3.4 136 US20030138772 SEQ ID NO: 54 AAVA3.4 137
US20030138772 SEQ ID NO: 68 AAVA3.5 138 US20030138772 SEQ ID NO: 55
AAVA3.5 139 US20030138772 SEQ ID NO: 69 AAVA3.7 140 US20030138772
SEQ ID NO: 56 AAVA3.7 141 US20030138772 SEQ ID NO: 67 AAV29.3 142
US20030138772 SEQ ID NO: 11 (AAVbb.1) AAVC2 143 US20030138772 SEQ
ID NO: 61 AAVCh.5 144 US20150159173 SEQ ID NO: 46, US20150315612
SEQ ID NO: 234 AAVcy.2 145 US20030138772 SEQ ID NO: 15 (AAV13.3)
AAV24.1 146 US20030138772 SEQ ID NO: 101 AAVcy.3 147 US20030138772
SEQ ID NO: 16 (AAV24.1) AAV27.3 148 US20030138772 SEQ ID NO: 104
AAVcy.4 149 US20030138772 SEQ ID NO: 17 (AAV27.3) AAVcy.5 150
US20150315612 SEQ ID NO: 227 AAV7.2 151 US20030138772 SEQ ID NO:
103 AAVcy.5 152 US20030138772 SEQ ID NO: 18 (AAV7.2) AAV16.3 153
US20030138772 SEQ ID NO: 105 AAVcy.6 154 US20030138772 SEQ ID NO:
10 (AAV16.3) AAVcy.5 155 US20150159173 SEQ ID NO: 8 AAVcy.5 156
US20150159173 SEQ ID NO: 24 AAVCy.5R1 157 US20150159173 AAVCy.5R2
158 US20150159173 AAVCy.5R3 159 US20150159173 AAVCy.5R4 160
US20150159173 AAVDJ 161 US20140359799 SEQ ID NO: 3, U.S. Pat. No.
7,588,772 SEQ ID NO: 2 AAVDJ 162 US20140359799 SEQ ID NO: 2, U.S.
Pat. No. 7,588,772 SEQ ID NO: 1 AAVDJ-8 163 U.S. Pat. No.
7,588,772; Grimm et al 2008 AAVDJ-8 164 U.S. Pat. No. 7,588,772;
Grimm et al 2008 AAVF5 165 US20030138772 SEQ ID NO: 110 AAVH2 166
US20030138772 SEQ ID NO: 26 AAVH6 167 US20030138772 SEQ ID NO: 25
AAVhE1.1 168 U.S. Pat. No. 9,233,131 SEQ ID NO: 44 AAVhEr1.14 169
U.S. Pat. No. 9,233,131 SEQ ID NO: 46 AAVhEr1.16 170 U.S. Pat. No.
9,233,131 SEQ ID NO: 48 AAVhEr1.18 171 U.S. Pat. No. 9,233,131 SEQ
ID NO: 49
AAVhEr1.23 172 U.S. Pat. No. 9,233,131 (AAVhEr2.29) SEQ ID NO: 53
AAVhEr1.35 173 U.S. Pat. No. 9,233,131 SEQ ID NO: 50 AAVhEr1.36 174
U.S. Pat. No. 9,233,131 SEQ ID NO: 52 AAVhEr1.5 175 U.S. Pat. No.
9,233,131 SEQ ID NO: 45 AAVhEr1.7 176 U.S. Pat. No. 9,233,131 SEQ
ID NO: 51 AAVhEr1.8 177 U.S. Pat. No. 9,233,131 SEQ ID NO: 47
AAVhEr2.16 178 U.S. Pat. No. 9,233,131 SEQ ID NO: 55 AAVhEr2.30 179
U.S. Pat. No. 9,233,131 SEQ ID NO: 56 AAVhEr2.31 180 U.S. Pat. No.
9,233,131 SEQ ID NO: 58 AAVhEr2.36 181 U.S. Pat. No. 9,233,131 SEQ
ID NO: 57 AAVhEr2.4 182 U.S. Pat. No. 9,233,131 SEQ ID NO: 54
AAVhEr3.1 183 U.S. Pat. No. 9,233,131 SEQ ID NO: 59 AAVhu.1 184
US20150315612 SEQ ID NO: 46 AAVhu.1 185 US20150315612 SEQ ID NO:
144 AAVhu.10 186 US20150315612 SEQ ID NO: 56 (AAV16.8) AAVhu.10 187
US20150315612 SEQ ID NO: 156 (AAV16.8) AAVhu.11 188 US20150315612
SEQ ID NO: 57 (AAV16.12) AAVhu.11 189 US20150315612 SEQ ID NO: 153
(AAV16.12) AAVhu.12 190 US20150315612 SEQ ID NO: 59 AAVhu.12 191
US20150315612 SEQ ID NO: 154 AAVhu.13 192 US20150159173 SEQ ID NO:
16, US20150315612 SEQ ID NO: 71 AAVhu.13 193 US20150159173 SEQ ID
NO: 32, US20150315612 SEQ ID NO: 129 AAVhu.136.1 194 US20150315612
SEQ ID NO: 165 AAVhu.140.1 195 US20150315612 SEQ ID NO: 166
AAVhu.140.2 196 US20150315612 SEQ ID NO: 167 AAVhu.145.6 197
US20150315612 SEQ ID No: 178 AAVhu.15 198 US20150315612 SEQ ID NO:
147 AAVhu.15 199 US20150315612 SEQ ID NO: 50 (AAV33.4) AAVhu.156.1
200 US20150315612 SEQ ID No: 179 AAVhu.16 201 US20150315612 SEQ ID
NO: 148 AAVhu.16 202 US20150315612 SEQ ID NO: 51 (AAV33.8) AAVhu.17
203 US20150315612 SEQ ID NO: 83 AAVhu.17 204 US20150315612 SEQ ID
NO: 4 (AAV33.12) AAVhu.172.1 205 US20150315612 SEQ ID NO: 171
AAVhu.172.2 206 US20150315612 SEQ ID NO: 172 AAVhu.173.4 207
US20150315612 SEQ ID NO: 173 AAVhu.173.8 208 US20150315612 SEQ ID
NO: 175 AAVhu.18 209 US20150315612 SEQ ID NO: 52 AAVhu.18 210
US20150315612 SEQ ID NO: 149 AAVhu.19 211 US20150315612 SEQ ID NO:
62 AAVhu.19 212 US20150315612 SEQ ID NO: 133 AAVhu.2 213
US20150315612 SEQ ID NO: 48 AAVhu.2 214 US20150315612 SEQ ID NO:
143 AAVhu.20 215 US20150315612 SEQ ID NO: 63 AAVhu.20 216
US20150315612 SEQ ID NO: 134 AAVhu.21 217 US20150315612 SEQ ID NO:
65 AAVhu.21 218 US20150315612 SEQ ID NO: 135 AAVhu.22 219
US20150315612 SEQ ID NO: 67 AAVhu.22 220 US20150315612 SEQ ID NO:
138 AAVhu.23 221 US20150315612 SEQ ID NO: 60 AAVhu.23.2 222
US20150315612 SEQ ID NO: 137 AAVhu.24 223 US20150315612 SEQ ID NO:
66 AAVhu.24 224 US20150315612 SEQ ID NO: 136 AAVhu.25 225
US20150315612 SEQ ID NO: 49 AAVhu.25 226 US20150315612 SEQ ID NO:
146 AAVhu.26 227 US20150159173 SEQ ID NO: 17, US20150315612 SEQ ID
NO: 61 AAVhu.26 228 US20150159173 SEQ ID NO: 33, US20150315612 SEQ
ID NO: 139 AAVhu.27 229 US20150315612 SEQ ID NO: 64 AAVhu.27 230
US20150315612 SEQ ID NO: 140 AAVhu.28 231 US20150315612 SEQ ID NO:
68 AAVhu.28 232 US20150315612 SEQ ID NO: 130 AAVhu.29 233
US20150315612 SEQ ID NO: 69 AAVhu.29 234 US20150159173 SEQ ID NO:
42, US20150315612 SEQ ID NO: 132 AAVhu.29 235 US20150315612 SEQ ID
NO: 225 AAVhu.29R 236 US20150159173 AAVhu.3 237 US20150315612 SEQ
ID NO: 44 AAVhu.3 238 US20150315612 SEQ ID NO: 145 AAVhu.30 239
US20150315612 SEQ ID NO: 70 AAVhu.30 240 US20150315612 SEQ ID NO:
131 AAVhu.31 241 US20150315612 SEQ ID NO: 1 AAVhu.31 242
US20150315612 SEQ ID NO: 121 AAVhu.32 243 US20150315612 SEQ ID NO:
2 AAVhu.32 244 US20150315612 SEQ ID NO: 122 AAVhu.33 245
US20150315612 SEQ ID NO: 75 AAVhu.33 246 US20150315612 SEQ ID NO:
124 AAVhu.34 247 US20150315612 SEQ ID NO: 72 AAVhu.34 248
US20150315612 SEQ ID NO: 125 AAVhu.35 249 US20150315612 SEQ ID NO:
73 AAVhu.35 250 US20150315612 SEQ ID NO: 164 AAVhu.36 251
US20150315612 SEQ ID NO: 74 AAVhu.36 252 US20150315612 SEQ ID NO:
126 AAVhu.37 253 US20150159173 SEQ ID NO: 34, US20150315612 SEQ ID
NO: 88 AAVhu.37 254 US20150315612 SEQ ID NO: 10, (AAV106.1)
US20150159173 SEQ ID NO: 18 AAVhu.38 255 US20150315612 SEQ ID NO:
161 AAVhu.39 256 US20150315612 SEQ ID NO: 102 AAVhu.39 257
US20150315612 SEQ ID NO: 24 (AAVLG-9) AAVhu.4 258 US20150315612 SEQ
ID NO: 47 AAVhu.4 259 US20150315612 SEQ ID NO: 141 AAVhu.40 260
US20150315612 SEQ ID NO: 87 AAVhu.40 261 US20150315612 SEQ ID No:
11 (AAV114.3) AAVhu.41 262 US20150315612 SEQ ID NO: 91 AAVhu.41 263
US20150315612 SEQ ID NO: 6 (AAV127.2) AAVhu.42 264 US20150315612
SEQ ID NO: 85 AAVhu.42 265 US20150315612 SEQ ID NO: 8 (AAV127.5)
AAVhu.43 266 US20150315612 SEQ ID NO: 160 AAVhu.43 267
US20150315612 SEQ ID NO: 236 AAVhu.43 268 US20150315612 SEQ ID NO:
80 (AAV128.1) AAVhu.44 269 US20150159173 SEQ ID NO: 45,
US20150315612 SEQ ID NO: 158 AAVhu.44 270 US20150315612 SEQ ID NO:
81 (AAV128.3) AAVhu.44R1 271 US20150159173 AAVhu.44R2 272
US20150159173 AAVhu.44R3 273 US20150159173 AAVhu.45 274
US20150315612 SEQ ID NO: 76 AAVhu.45 275 US20150315612 SEQ ID NO:
127 AAVhu.46 276 US20150315612 SEQ ID NO: 82 AAVhu.46 277
US20150315612 SEQ ID NO: 159 AAVhu.46 278 US20150315612 SEQ ID NO:
224 AAVhu.47 279 US20150315612 SEQ ID NO: 77 AAVhu.47 280
US20150315612 SEQ ID NO: 128 AAVhu.48 281 US20150159173 SEQ ID NO:
38 AAVhu.48 282 US20150315612 SEQ ID NO: 157 AAVhu.48 283
US20150315612 SEQ ID NO: 78 (AAV130.4) AAVhu.48R1 284 US20150159173
AAVhu.48R2 285 US20150159173 AAVhu.48R3 286 US20150159173 AAVhu.49
287 US20150315612 SEQ ID NO: 209 AAVhu.49 288 US20150315612 SEQ ID
NO: 189 AAVhu.5 289 US20150315612 SEQ ID NO: 45 AAVhu.5 290
US20150315612 SEQ ID NO: 142 AAVhu.51 291 US20150315612 SEQ ID NO:
208 AAVhu.51 292 US20150315612 SEQ ID NO: 190 AAVhu.52 293
US20150315612 SEQ ID NO: 210 AAVhu.52 294 US20150315612 SEQ ID NO:
191 AAVhu.53 295 US20150159173 SEQ ID NO: 19 AAVhu.53 296
US20150159173 SEQ ID NO: 35 AAVhu.53 297 US20150315612 SEQ ID NO:
176 (AAV145.1) AAVhu.54 298 US20150315612 SEQ ID NO: 188 AAVhu.54
299 US20150315612 SEQ ID No: 177 (AAV145.5) AAVhu.55 300
US20150315612 SEQ ID NO: 187 AAVhu.56 301 US20150315612 SEQ ID NO:
205 AAVhu.56 302 US20150315612 SEQ ID NO: 168 (AAV145.6) AAVhu.56
303 US20150315612 SEQ ID NO: 192 (AAV145.6) AAVhu.57 304
US20150315612 SEQ ID NO: 206 AAVhu.57 305 US20150315612 SEQ ID NO:
169 AAVhu.57 306 US20150315612 SEQ ID NO: 193 AAVhu.58 307
US20150315612 SEQ ID NO: 207 AAVhu.58 308 US20150315612 SEQ ID NO:
194 AAVhu.6 309 US20150315612 SEQ ID NO: 5 (AAV3.1) AAVhu.6 310
US20150315612 SEQ ID NO: 84 (AAV3.1) AAVhu.60 311 US20150315612 SEQ
ID NO: 184 AAVhu.60 312 US20150315612 SEQ ID NO: 170 (AAV161.10)
AAVhu.61 313 US20150315612 SEQ ID NO: 185 AAVhu.61 314
US20150315612 SEQ ID NO: 174 (AAV161.6) AAVhu.63 315 US20150315612
SEQ ID NO: 204 AAVhu.63 316 US20150315612 SEQ ID NO: 195 AAVhu.64
317 US20150315612 SEQ ID NO: 212 AAVhu.64 318 US20150315612 SEQ ID
NO: 196 AAVhu.66 319 US20150315612 SEQ ID NO: 197 AAVhu.67 320
US20150315612 SEQ ID NO: 215 AAVhu.67 321 US20150315612 SEQ ID NO:
198 AAVhu.7 322 US20150315612 SEQ ID NO: 226 AAVhu.7 323
US20150315612 SEQ ID NO: 150 AAVhu.7 324 US20150315612 SEQ ID NO:
55 (AAV7.3) AAVhu.71 325 US20150315612 SEQ ID NO: 79 AAVhu.8 326
US20150315612 SEQ ID NO: 53 AAVhu.8 327 US20150315612 SEQ ID NO: 12
AAVhu.8 328 US20150315612 SEQ ID NO: 151 AAVhu.9 329 US20150315612
SEQ ID NO: 58 (AAV3.1) AAVhu.9 330 US20150315612 SEQ ID NO: 155
(AAV3.1) AAV-LK01 331 US20150376607 SEQ ID NO: 2 AAV-LK01 332
US20150376607 SEQ ID NO: 29 AAV-LK02 333 US20150376607 SEQ ID NO: 3
AAV-LK02 334 US20150376607 SEQ ID NO: 30 AAV-LK03 335 US20150376607
SEQ ID NO: 4 AAV-LK03 336 WO2015121501 SEQ ID NO: 12, US20150376607
SEQ ID NO: 31 AAV-LK04 337 US20150376607 SEQ ID NO: 5 AAV-LK04 338
US20150376607 SEQ ID NO: 32 AAV-LK05 339 US20150376607 SEQ ID NO: 6
AAV-LK05 340 US20150376607 SEQ ID NO: 33 AAV-LK06 341 US20150376607
SEQ ID NO: 7 AAV-LK06 342 US20150376607 SEQ ID NO: 34 AAV-LK07 343
US20150376607 SEQ ID NO: 8 AAV-LK07 344 US20150376607 SEQ ID NO: 35
AAV-LK08 345 US20150376607 SEQ ID NO: 9 AAV-LK08 346 US20150376607
SEQ ID NO: 36 AAV-LK09 347 US20150376607 SEQ ID NO: 10 AAV-LK09 348
US20150376607 SEQ ID NO: 37 AAV-LK10 349 US20150376607 SEQ ID NO:
11 AAV-LK10 350 US20150376607 SEQ ID NO: 38 AAV-LK11 351
US20150376607 SEQ ID NO: 12 AAV-LK11 352 US20150376607 SEQ ID NO:
39 AAV-LK12 353 US20150376607 SEQ ID NO: 13 AAV-LK12 354
US20150376607 SEQ ID NO: 40 AAV-LK13 355 US20150376607 SEQ ID NO:
14 AAV-LK13 356 US20150376607 SEQ ID NO: 41 AAV-LK14 357
US20150376607 SEQ ID NO: 15 AAV-LK14 358 US20150376607 SEQ ID NO:
42 AAV-LK15 359 US20150376607 SEQ ID NO: 16 AAV-LK15 360
US20150376607 SEQ ID NO: 43 AAV-LK16 361 US20150376607 SEQ ID NO:
17 AAV-LK16 362 US20150376607 SEQ ID NO: 44 AAV-LK17 363
US20150376607 SEQ ID NO: 18 AAV-LK17 364 US20150376607 SEQ ID NO:
45 AAV-LK18 365 US20150376607 SEQ ID NO: 19 AAV-LK18 366
US20150376607 SEQ ID NO: 46 AAV-LK19 367 US20150376607 SEQ ID NO:
20 AAV-LK19 368 US20150376607 SEQ ID NO: 47 AAV-PAEC 369
US20150376607 SEQ ID NO: 1 AAV-PAEC 370 US20150376607 SEQ ID NO: 48
AAV-PAEC11 371 US20150376607 SEQ ID NO: 26 AAV-PAEC11 372
US20150376607 SEQ ID NO: 54 AAV-PAEC12 373 US20150376607 SEQ ID NO:
27 AAV-PAEC12 374 US20150376607 SEQ ID NO: 51 AAV-PAEC13 375
US20150376607 SEQ ID NO: 28 AAV-PAEC13 376 US20150376607 SEQ ID NO:
49
AAV-PAEC2 377 US20150376607 SEQ ID NO: 21 AAV-PAEC2 378
US20150376607 SEQ ID NO: 56 AAV-PAEC4 379 US20150376607 SEQ ID NO:
22 AAV-PAEC4 380 US20150376607 SEQ ID NO: 55 AAV-PAEC6 381
US20150376607 SEQ ID NO: 23 AAV-PAEC6 382 US20150376607 SEQ ID NO:
52 AAV-PAEC7 383 US20150376607 SEQ ID NO: 24 AAV-PAEC7 384
US20150376607 SEQ ID NO: 53 AAV-PAEC8 385 US20150376607 SEQ ID NO:
25 AAV-PAEC8 386 US20150376607 SEQ ID NO: 50 AAVpi.1 387
US20150315612 SEQ ID NO: 28 AAVpi.1 388 US20150315612 SEQ ID NO: 93
AAVpi.2 389 US20150315612 SEQ ID NO: 30 AAVpi.2 390 US20150315612
SEQ ID NO: 95 AAVpi.3 391 US20150315612 SEQ ID NO: 29 AAVpi.3 392
US20150315612 SEQ ID NO: 94 AAVrh.10 393 US20150159173 SEQ ID NO: 9
AAVrh.10 394 US20150159173 SEQ ID NO: 25 AAV44.2 395 US20030138772
SEQ ID NO: 59 AAVrh.10 396 US20030138772 SEQ ID NO: 81 (AAV44.2)
AAV42.1B 397 US20030138772 SEQ ID NO: 90 AAVrh.12 398 US20030138772
SEQ ID NO: 30 (AAV42.1b) AAVrh.13 399 US20150159173 SEQ ID NO: 10
AAVrh.13 400 US20150159173 SEQ ID NO: 26 AAVrh.13 401 US20150315612
SEQ ID NO: 228 AAVrh.13R 402 US20150159173 AAV42.3A 403
US20030138772 SEQ ID NO: 87 AAVrh.14 404 US20030138772 SEQ ID NO:
32 (AAV42.3a) AAV42.5A 405 US20030138772 SEQ ID NO: 89 AAVrh.17 406
US20030138772 SEQ ID NO: 34 (AAV42.5a) AAV42.5B 407 US20030138772
SEQ ID NO: 91 AAVrh.18 408 US20030138772 SEQ ID NO: 29 (AAV42.5b)
AAV42.6B 409 US20030138772 SEQ ID NO: 112 AAVrh.19 410
US20030138772 SEQ ID NO: 38 (AAV42.6b) AAVrh.2 411 US20150159173
SEQ ID NO: 39 AAVrh.2 412 US20150315612 SEQ ID NO: 231 AAVrh.20 413
US20150159173 SEQ ID NO: 1 AAV42.10 414 US20030138772 SEQ ID NO:
106 AAVrh.21 415 US20030138772 SEQ ID NO: 35 (AAV42.10) AAV42.11
416 US20030138772 SEQ ID NO: 108 AAVrh.22 417 US20030138772 SEQ ID
NO: 37 (AAV42.11) AAV42.12 418 US20030138772 SEQ ID NO: 113
AAVrh.23 419 US20030138772 SEQ ID NO: 58 (AAV42.12) AAV42.13 420
US20030138772 SEQ ID NO: 86 AAVrh.24 421 US20030138772 SEQ ID NO:
31 (AAV42.13) AAV42.15 422 US20030138772 SEQ ID NO: 84 AAVrh.25 423
US20030138772 SEQ ID NO: 28 (AAV42.15) AAVrh.2R 424 US20150159173
AAVrh.31 425 US20030138772 SEQ ID NO: 48 (AAV223.1) AAVC1 426
US20030138772 SEQ ID NO: 60 AAVrh.32 427 US20030138772 SEQ ID NO:
19 (AAVC1) AAVrh.32/33 428 US20150159173 SEQ ID NO: 2 AAVrh.33 429
US20030138772 SEQ ID NO: 20 (AAVC3) AAVC5 430 US20030138772 SEQ ID
NO: 62 AAVrh.34 431 US20030138772 SEQ ID NO: 21 (AAVC5) AAVF1 432
US20030138772 SEQ ID NO: 109 AAVrh.35 433 US20030138772 SEQ ID NO:
22 (AAVF1) AAVF3 434 US20030138772 SEQ ID NO: 111 AAVrh.36 435
US20030138772 SEQ ID NO: 23 (AAVF3) AAVrh.37 436 US20030138772 SEQ
ID NO: 24 AAVrh.37 437 US20150159173 SEQ ID NO: 40 AAVrh.37 438
US20150315612 SEQ ID NO: 229 AAVrh.37R2 439 US20150159173 AAVrh.38
440 US20150315612 SEQ ID NO: 7 (AAVLG-4) AAVrh.38 441 US20150315612
SEQ ID NO: 86 (AAVLG-4) AAVrh.39 442 US20150159173 SEQ ID NO: 20,
US20150315612 SEQ ID NO: 13 AAVrh.39 443 US20150159173 SEQ ID NO:
3, US20150159173 SEQ ID NO: 36, US20150315612 SEQ ID NO: 89
AAVrh.40 444 US20150315612 SEQ ID NO: 92 AAVrh.40 445 US20150315612
SEQ ID No: 14 (AAVLG-10) AAVrh.43 446 US20150315612 SEQ ID NO: 43,
(AAVN721-8) US20150159173 SEQ ID NO: 21 AAVrh.43 447 US20150315612
SEQ ID NO: 163, (AAVN721-8) US20150159173 SEQ ID NO: 37 AAVrh.44
448 US20150315612 SEQ ID NO: 34 AAVrh.44 449 US20150315612 SEQ ID
NO: 111 AAVrh.45 450 US20150315612 SEQ ID NO: 41 AAVrh.45 451
US20150315612 SEQ ID NO: 109 AAVrh.46 452 US20150159173 SEQ ID NO:
22, US20150315612 SEQ ID NO: 19 AAVrh.46 453 US20150159173 SEQ ID
NO: 4, US20150315612 SEQ ID NO: 101 AAVrh.47 454 US20150315612 SEQ
ID NO: 38 AAVrh.47 455 US20150315612 SEQ ID NO: 118 AAVrh.48 456
US20150159173 SEQ ID NO: 44, US20150315612 SEQ ID NO: 115
AAVrh.48.1 457 US20150159173 AAVrh.48.1.2 458 US20150159173
AAVrh.48.2 459 US20150159173 AAVrh.48 460 US20150315612 SEQ ID NO:
32 (AAV1-7) AAVrh.49 461 US20150315612 SEQ ID NO: 25 (AAV1-8)
AAVrh.49 462 US20150315612 SEQ ID NO: 103 (AAV1-8) AAVrh.50 463
US20150315612 SEQ ID NO: 23 (AAV2-4) AAVrh.50 464 US20150315612 SEQ
ID NO: 108 (AAV2-4) AAVrh.51 465 US20150315612 SEQ ID No: 22
(AAV2-5) AAVrh.51 466 US20150315612 SEQ ID NO: 104 (AAV2-5)
AAVrh.52 467 US20150315612 SEQ ID NO: 18 (AAV3-9) AAVrh.52 468
US20150315612 SEQ ID NO: 96 (AAV3-9) AAVrh.53 469 US20150315612 SEQ
ID NO: 97 AAVrh.53 470 US20150315612 SEQ ID NO: 17 (AAV3-11)
AAVrh.53 471 US20150315612 SEQ ID NO: 186 (AAV3-11) AAVrh.54 472
US20150315612 SEQ ID NO: 40 AAVrh.54 473 US20150159173 SEQ ID NO:
49, US20150315612 SEQ ID NO: 116 AVrh.55 474 US20150315612 SEQ ID
NO: 37 AAVrh.55 475 US20150315612 SEQ ID NO: 117 (AAV4-19) AAVrh.56
476 US20150315612 SEQ ID NO: 54 AAVrh.56 477 US20150315612 SEQ ID
NO: 152 AAVrh.57 478 US20150315612 SEQ ID NO: 26 AAVrh.57 479
US20150315612 SEQ ID NO: 105 AAVrh.58 480 US20150315612 SEQ ID NO:
27 AAVrh.58 481 US20150159173 SEQ ID NO: 48, US20150315612 SEQ ID
NO: 106 AAVrh.58 482 US20150315612 SEQ ID NO: 232 AAVrh.59 483
US20150315612 SEQ ID NO: 42 AAVrh.59 484 US20150315612 SEQ ID NO:
110 AAVrh.60 485 US20150315612 SEQ ID NO: 31 AAVrh.60 486
US20150315612 SEQ ID NO: 120 AAVrh.61 487 US20150315612 SEQ ID NO:
107 AAVrh.61 488 US20150315612 SEQ ID NO: 21 (AAV2-3) AAVrh.62 489
US20150315612 SEQ ID No: 33 (AAV2-15) AAVrh.62 490 US20150315612
SEQ ID NO: 114 (AAV2-15) AAVrh.64 491 US20150315612 SEQ ID No: 15
AAVrh.64 492 US20150159173 SEQ ID NO: 43, US20150315612 SEQ ID NO:
99 AAVrh.64 493 US20150315612 SEQ ID NO: 233 AAVRh.64R1 494
US20150159173 AAVRh.64R2 495 US20150159173 AAVrh.65 496
US20150315612 SEQ ID NO: 35 AAVrh.65 497 US20150315612 SEQ ID NO:
112 AAVrh.67 498 US20150315612 SEQ ID NO: 36 AAVrh.67 499
US20150315612 SEQ ID NO: 230 AAVrh.67 500 US20150159173 SEQ ID NO:
47, US20150315612 SEQ ID NO: 113 AAVrh.68 501 US20150315612 SEQ ID
NO: 16 AAVrh.68 502 US20150315612 SEQ ID NO: 100 AAVrh.69 503
US20150315612 SEQ ID NO: 39 AAVrh.69 504 US20150315612 SEQ ID NO:
119 AAVrh.70 505 US20150315612 SEQ ID NO: 20 AAVrh.70 506
US20150315612 SEQ ID NO: 98 AAVrh.71 507 US20150315612 SEQ ID NO:
162 AAVrh.72 508 US20150315612 SEQ ID NO: 9 AAVrh.73 509
US20150159173 SEQ ID NO: 5 AAVrh.74 510 US20150159173 SEQ ID NO: 6
AAVrh.8 511 US20150159173 SEQ ID NO: 41 AAVrh.8 512 US20150315612
SEQ ID NO: 235 AAVrh.8R 513 US20150159173, WO2015168666 SEQ ID NO:
9 AAVrh.8R 514 WO2015168666 SEQ ID NO: 10 A586R mutant AAVrh.8R 515
WO2015168666 SEQ ID NO: 11 R533A mutant BAAV 516 U.S. Pat. No.
9,193,769 (bovine AAV) SEQ ID NO: 8 BAAV 517 U.S. Pat. No.
9,193,769 (bovine AAV) SEQ ID NO: 10 BAAV 518 U.S. Pat. No.
9,193,769 (bovine AAV) SEQ ID NO: 4 BAAV 519 U.S. Pat. No.
9,193,769 (bovine AAV) SEQ ID NO: 2 BAAV 520 U.S. Pat. No.
9,193,769 (bovine AAV) SEQ ID NO: 6 BAAV 521 U.S. Pat. No.
9,193,769 (bovine AAV) SEQ ID NO: 1 BAAV 522 U.S. Pat. No.
9,193,769 (bovine AAV) SEQ ID NO: 5 BAAV 523 U.S. Pat. No.
9,193,769 (bovine AAV) SEQ ID NO: 3 BAAV 524 U.S. Pat. No.
9,193,769 (bovine AAV) SEQ ID NO: 11 BAAV 525 U.S. Pat. No.
7,427,396 (bovine AAV) SEQ ID NO: 5 BAAV 526 U.S. Pat. No.
7,427,396 (bovine AAV) SEQ ID NO: 6 BAAV 527 U.S. Pat. No.
9,193,769 (bovine AAV) SEQ ID NO: 7 BAAV 528 U.S. Pat. No.
9,193,769 (bovine AAV) SEQ ID NO: 9 BNP61 AAV 529 US20150238550 SEQ
ID NO: 1 BNP61 AAV 530 US20150238550 SEQ ID NO: 2 BNP62 AAV 531
US20150238550 SEQ ID NO: 3 BNP63 AAV 532 US20150238550 SEQ ID NO: 4
caprine AAV 533 U.S. Pat. No. 7,427,396 SEQ ID NO: 3 caprine AAV
534 U.S. Pat. No. 7,4273,96 SEQ ID NO: 4 true type AAV 535
WO2015121501 SEQ ID NO: 2 (ttAAV) AAAV 536 U.S. Pat. No. 9,238,800
(Avian AAV) SEQ ID NO: 12 AAAV 537 U.S. Pat. No. 9,238,800 (Avian
AAV) SEQ ID NO: 2 AAAV 538 U.S. Pat. No. 9,238,800 (Avian AAV) SEQ
ID NO: 6 AAAV 539 U.S. Pat. No. 9,238,800 (Avian AAV) SEQ ID NO: 4
AAAV 540 U.S. Pat. No. 9,238,800 (Avian AAV) SEQ ID NO: 8 AAAV 541
U.S. Pat. No. 9,238,800 (Avian AAV) SEQ ID NO: 14 AAAV 542 U.S.
Pat. No. 9238,800 (Avian AAV) SEQ ID NO: 10 AAAV 543 U.S. Pat. No.
9,238,800 (Avian AAV) SEQ ID NO: 15 AAAV 544 U.S. Pat. No.
9,238,800 (Avian AAV) SEQ ID NO: 5 AAAV 545 U.S. Pat. No. 9,238,800
(Avian AAV) SEQ ID NO: 9 AAAV 546 U.S. Pat. No. 9,238,800 (Avian
AAV) SEQ ID NO: 3 AAAV 547 U.S. Pat. No. 9,238,800 (Avian AAV) SEQ
ID NO: 7 AAAV 548 U.S. Pat. No. 9,238,800 (Avian AAV) SEQ ID NO:
11
AAAV 549 U.S. Pat. No. 9,238,800 (Avian AAV) SEQ ID NO: 13 AAAV 550
U.S. Pat. No. 9,238,800 (Avian AAV) SEQ ID NO: 1 AAV Shuffle 551
US20160017295 SEQ ID NO: 23 100-1 AAV Shuffle 552 US20160017295 SEQ
ID NO: 11 100-1 AAV Shuffle 553 US20160017295 SEQ ID NO: 37 100-2
AAV Shuffle 554 US20160017295 SEQ ID NO: 29 100-2 AAV Shuffle 555
US20160017295 SEQ ID NO: 24 100-3 AAV Shuffle 556 US20160017295 SEQ
ID NO: 12 100-3 AAV Shuffle 557 US20160017295 SEQ ID NO: 25 100-7
AAV Shuffle 558 US20160017295 SEQ ID NO: 13 100-7 AAV Shuffle 559
US20160017295 SEQ ID NO: 34 10-2 AAV Shuffle 560 US20160017295 SEQ
ID NO: 26 10-2 AAV Shuffle 561 US20160017295 SEQ ID NO: 35 10-6 AAV
Shuffle 562 US20160017295 SEQ ID NO: 27 10-6 AAV Shuffle 563
US20160017295 SEQ ID NO: 36 10-8 AAV Shuffle 564 US20160017295 SEQ
ID NO: 28 10-8 AAV SM 100-10 565 US20160017295 SEQ ID NO: 41 AAV SM
100-10 566 US20160017295 SEQ ID NO: 33 AAV SM 100-3 567
US20160017295 SEQ ID NO: 40 AAV SM 100-3 568 US20160017295 SEQ ID
NO: 32 AAV SM 10-1 569 US20160017295 SEQ ID NO: 38 AAV SM 10-1 570
US20160017295 SEQ ID NO: 30 AAV SM 10-2 571 US20160017295 SEQ ID
NO: 10 AAV SM 10-2 572 US20160017295 SEQ ID NO: 22 AAV SM 10-8 573
US20160017295 SEQ ID NO: 39 AAV SM 10-8 574 US20160017295 SEQ ID
NO: 31 AAVF1/HSC1 575 WO2016049230 SEQ ID NO: 20 AAVF2/HSC2 576
WO2016049230 SEQ ID NO: 21 AAVF3/HSC3 577 WO2016049230 SEQ ID NO:
22 AAVF4/HSC4 578 WO2016049230 SEQ ID NO: 23 AAVF5/HSC5 579
WO2016049230 SEQ ID NO: 25 AAVF6/HSC6 580 WO2016049230 SEQ ID NO:
24 AAVF7/HSC7 581 WO2016049230 SEQ ID NO: 27 AAVF8/HSC8 582
WO2016049230 SEQ ID NO: 28 AAVF9/HSC9 583 WO2016049230 SEQ ID NO:
29 AAVF11/HSC11 584 WO2016049230 SEQ ID NO: 26 AAVF12/HSC12 585
WO2016049230 SEQ ID NO: 30 AAVF13/HSC13 586 WO2016049230 SEQ ID NO:
31 AAVF14/HSC14 587 WO2016049230 SEQ ID NO: 32 AAVF15/HSC15 588
WO2016049230 SEQ ID NO: 33 AAVF16/HSC16 589 WO2016049230 SEQ ID NO:
34 AAVF17/HSC17 590 WO2016049230 SEQ ID NO: 35 AAVF1/HSC1 591
WO2016049230 SEQ ID NO: 2 AAVF2/HSC2 592 WO2016049230 SEQ ID NO: 3
AAVF3/HSC3 593 WO2016049230 SEQ ID NO: 5 AAVF4/HSC4 594
WO2016049230 SEQ ID NO: 6 AAVF5/HSC5 595 WO2016049230 SEQ ID NO: 11
AAVF6/HSC6 596 WO2016049230 SEQ ID NO: 7 AAVF7/HSC7 597
WO2016049230 SEQ ID NO: 8 AAVF8/HSC8 598 WO2016049230 SEQ ID NO: 9
AAVF9/HSC9 599 WO2016049230 SEQ ID NO: 10 AAVF11/HSC11 600
WO2016049230 SEQ ID NO: 4 AAVF12/HSC12 601 WO2016049230 SEQ ID NO:
12 AAVF13/HSC13 602 WO2016049230 SEQ ID NO: 14 AAVF14/HSC14 603
WO2016049230 SEQ ID NO: 15 AAVF15/HSC15 604 WO2016049230 SEQ ID NO:
16 AAVF16/HSC16 605 WO2016049230 SEQ ID NO: 17 AAVF17/HSC17 606
WO2016049230 SEQ ID NO: 13 AAV CBr-E1 607 U.S. Pat. No. 8,734,809
SEQ ID NO: 13 AAV CBr-E2 608 U.S. Pat. No. 8,734,809 SEQ ID NO: 14
AAV CBr-E3 609 U.S. Pat. No. 8,734,809 SEQ ID NO: 15 AAV CBr-E4 610
U.S. Pat. No. 8,734,809 SEQ ID NO: 16 AAV CBr-E5 611 U.S. Pat. No.
8,734,809 SEQ ID NO: 17 AAV CBr-e5 612 U.S. Pat. No. 8,734,809 SEQ
ID NO: 18 AAV CBr-E6 613 U.S. Pat. No. 8,734,809 SEQ ID NO: 19 AAV
CBr-E7 614 U.S. Pat. No. 8,734,809 SEQ ID NO: 20 AAV CBr-E8 615
U.S. Pat. No. 8,734,809 SEQ ID NO: 21 AAV CLv-D1 616 U.S. Pat. No.
8,73,4809 SEQ ID NO: 22 AAV CLv-D2 617 U.S. Pat. No. 8,734,809 SEQ
ID NO: 23 AAV CLv-D3 618 U.S. Pat. No. 8,734,809 SEQ ID NO: 24 AAV
CLv-D4 619 U.S. Pat. No. 8,734,809 SEQ ID NO: 25 AAV CLv-D5 620
U.S. Pat. No. 8,734,809 SEQ ID NO: 26 AAV CLv-D6 621 U.S. Pat. No.
8,734,809 SEQ ID NO: 27 AAV CLv-D7 622 U.S. Pat. No. 8,734,809 SEQ
ID NO: 28 AAV CLv-D8 623 U.S. Pat. No. 8,734,809 SEQ ID NO: 29 AAV
CLv-E1 624 U.S. Pat. No. 8,734,809 SEQ ID NO: 13 AAV CLv-R1 625
U.S. Pat. No. 8,734,809 SEQ ID NO: 30 AAV CLv-R2 626 U.S. Pat. No.
8,734,809 SEQ ID NO: 31 AAV CLv-R3 627 U.S. Pat. No. 8,734,809 SEQ
ID NO: 32 AAV CLv-R4 628 U.S. Pat. No. 8,734,809 SEQ ID NO: 33 AAV
CLv-R5 629 U.S. Pat. No. 8,734,809 SEQ ID NO: 34 AAV CLv-R6 630
U.S. Pat. No. 8,734,809 SEQ ID NO: 35 AAV CLv-R7 631 U.S. Pat. No.
8,734,809 SEQ ID NO: 36 AAV CLv-R8 632 U.S. Pat. No. 8,734,809 SEQ
ID NO: 37 AAV CLv-R9 633 U.S. Pat. No. 8,734,809 SEQ ID NO: 38 AAV
CLg-F1 634 U.S. Pat. No. 8,734,809 SEQ ID NO: 39 AAV CLg-F2 635
U.S. Pat. No. 8,734,809 SEQ ID NO: 40 AAV CLg-F3 636 U.S. Pat. No.
8,734,809 SEQ ID NO: 41 AAV CLg-F4 637 U.S. Pat. No. 8,734,809 SEQ
ID NO: 42 AAV CLg-F5 638 U.S. Pat. No. 8,734,809 SEQ ID NO: 43 AAV
CLg-F6 639 U.S. Pat. No. 8,734,809 SEQ ID NO: 43 AAV CLg-F7 640
U.S. Pat. No. 8,734,809 SEQ ID NO: 44 AAV CLg-F8 641 U.S. Pat. No.
8,734,809 SEQ ID NO: 43 AAV CSp-1 642 U.S. Pat. No. 8,734,809 SEQ
ID NO: 45 AAV CSp-10 643 U.S. Pat. No. 8,734,809 SEQ ID NO: 46 AAV
CSp-11 644 U.S. Pat. No. 8,734,809 SEQ ID NO: 47 AAV CSp-2 645 U.S.
Pat. No. 8,734,809 SEQ ID NO: 48 AAV CSp-3 646 U.S. Pat. No.
8,734,809 SEQ ID NO: 49 AAV CSp-4 647 U.S. Pat. No. 8,734,809 SEQ
ID NO: 50 AAV CSp-6 648 U.S. Pat. No. 8,734,809 SEQ ID NO: 51 AAV
CSp-7 649 U.S. Pat. No. 8,734,809 SEQ ID NO: 52 AAV CSp-8 650 U.S.
Pat. No. 8,734,809 SEQ ID NO: 53 AAV CSp-9 651 U.S. Pat. No.
8,734,809 SEQ ID NO: 54 AAV CHt-2 652 U.S. Pat. No. 8,734,809 SEQ
ID NO: 55 AAV CHt-3 653 U.S. Pat. No. 8,734,809 SEQ ID NO: 56 AAV
CKd-1 654 U.S. Pat. No. 8,734,809 SEQ ID NO: 57 AAV CKd-10 655 U.S.
Pat. No. 8,734,809 SEQ ID NO: 58 AAV CKd-2 656 U.S. Pat. No.
8,734,809 SEQ ID NO: 59 AAV CKd-3 657 U.S. Pat. No. 8,734,809 SEQ
ID NO: 60 AAV CKd-4 658 U.S. Pat. No. 8,734,809 SEQ ID NO: 61 AAV
CKd-6 659 U.S. Pat. No. 8,734,809 SEQ ID NO: 62 AAV CKd-7 660 U.S.
Pat. No. 8,734,809 SEQ ID NO: 63 AAV CKd-8 661 U.S. Pat. No.
8,734,809 SEQ ID NO: 64 AAV CLv-1 662 U.S. Pat. No. 8,734,809 SEQ
ID NO: 65 AAV CLv-12 663 U.S. Pat. No. 8,734,809 SEQ ID NO: 66 AAV
CLv-13 664 U.S. Pat. No. 8,734,809 SEQ ID NO: 67 AAV CLv-2 665 U.S.
Pat. No. 8,734,809 SEQ ID NO: 68 AAV CLv-3 666 U.S. Pat. No.
8,734,809 SEQ ID NO: 69 AAV CLv-4 667 U.S. Pat. No. 8,734,809 SEQ
ID NO: 70 AAV CLv-6 668 U.S. Pat. No. 8,734,809 SEQ ID NO: 71 AAV
CLv-8 669 U.S. Pat. No. 8,734,809 SEQ ID NO: 72 AAV CKd-B1 670 U.S.
Pat. No. 8,734,809 SEQ ID NO: 73 AAV CKd-B2 671 U.S. Pat. No.
8,734,809 SEQ ID NO: 74 AAV CKd-B3 672 U.S. Pat. No. 8,734,809 SEQ
ID NO: 75 AAV CKd-B4 673 U.S. Pat. No. 8,734,809 SEQ ID NO: 76 AAV
CKd-B5 674 U.S. Pat. No. 8,734,809 SEQ ID NO: 77 AAV CKd-B6 675
U.S. Pat. No. 8,734,809 SEQ ID NO: 78 AAV CKd-B7 676 U.S. Pat. No.
8,734,809 SEQ ID NO: 79 AAV CKd-B8 677 U.S. Pat. No. 8,734,809 SEQ
ID NO: 80 AAV CKd-H1 678 U.S. Pat. No. 8,734,809 SEQ ID NO: 81 AAV
CKd-H2 679 U.S. Pat. No. 8,734,809 SEQ ID NO: 82 AAV CKd-H3 680
U.S. Pat. No. 8,734,809 SEQ ID NO: 83 AAV CKd-H4 681 U.S. Pat. No.
8,734,809 SEQ ID NO: 84 AAV CKd-H5 682 U.S. Pat. No. 8,734,809 SEQ
ID NO: 85 AAV CKd-H6 683 U.S. Pat. No. 8,734,809 SEQ ID NO: 77 AAV
CHt-1 684 U.S. Pat. No. 8,734,809 SEQ ID NO: 86 AAV CLv1-1 685 U.S.
Pat. No. 8,734,809 SEQ ID NO: 171 AAV CLv1-2 686 U.S. Pat. No.
8,734,809 SEQ ID NO: 172 AAV CLv1-3 687 U.S. Pat. No. 8,734,809 SEQ
ID NO: 173 AAV CLv1-4 688 U.S. Pat. No. 8,734,809 SEQ ID NO: 174
AAV C1v1-7 689 U.S. Pat. No. 8,734,809 SEQ ID NO: 175 AAV C1v1-8
690 U.S. Pat. No. 8,734,809 SEQ ID NO: 176 AAV Clv1-9 691 U.S. Pat.
No. 8,734,809 SEQ ID NO: 177 AAV Clv1-10 692 U.S. Pat. No.
8,734,809 SEQ ID NO: 178 AAV.VR-355 693 U.S. Pat. No. 8,734,809 SEQ
ID NO: 181 AAV.hu.48R3 694 U.S. Pat. No. 8,734,809 SEQ ID NO: 183
AAV CBr-E1 695 U.S. Pat. No. 8,734,809
SEQ ID NO: 87 AAV CBr-E2 696 U.S. Pat. No. 8,734,809 SEQ ID NO: 88
AAV CBr-E3 697 U.S. Pat. No. 8,734,809 SEQ ID NO: 89 AAV CBr-E4 698
U.S. Pat. No. 8,734,809 SEQ ID NO: 90 AAV CBr-E5 699 U.S. Pat. No.
8,734,809 SEQ ID NO: 91 AAV CBr-e5 700 U.S. Pat. No. 8,734,809 SEQ
ID NO: 92 AAV CBr-E6 701 U.S. Pat. No. 8,734,809 SEQ ID NO: 93 AAV
CBr-E7 702 U.S. Pat. No. 8,734,809 SEQ ID NO: 94 AAV CBr-E8 703
U.S. Pat. No. 8,734,809 SEQ ID NO: 95 AAV CLv-D1 704 U.S. Pat. No.
8,734,809 SEQ ID NO: 96 AAV CLv-D2 705 U.S. Pat. No. 8,734,809 SEQ
ID NO: 97 AAV CLv-D3 706 U.S. Pat. No. 8,734,809 SEQ ID NO: 98 AAV
CLv-D4 707 U.S. Pat. No. 8,734,809 SEQ ID NO: 99 AAV CLv-D5 708
U.S. Pat. No. 8,734,809 SEQ ID NO: 100 AAV CLv-D6 709 U.S. Pat. No.
8,734,809 SEQ ID NO: 101 AAV CLv-D7 710 U.S. Pat. No. 8,734,809 SEQ
ID NO: 102 AAV CLv-D8 711 U.S. Pat. No. 8,734,809 SEQ ID NO: 103
AAV CLv-E1 712 U.S. Pat. No. 8,734,809 SEQ ID NO: 87 AAV CLv-R1 713
U.S. Pat. No. 8,734,809 SEQ ID NO: 104 AAV CLv-R2 714 U.S. Pat. No.
8,734,809 SEQ ID NO: 105 AAV CLv-R3 715 U.S. Pat. No. 8,734,809 SEQ
ID NO: 106 AAV CLv-R4 716 U.S. Pat. No. 8,734,809 SEQ ID NO: 107
AAV CLv-R5 717 U.S. Pat. No. 8,734,809 SEQ ID NO: 108 AAV CLv-R6
718 U.S. Pat. No. 8,734,809 SEQ ID NO: 109 AAV CLv-R7 719 U.S. Pat.
No. 8,734,809 SEQ ID NO: 110 AAV CLv-R8 720 U.S. Pat. No. 8,734,809
SEQ ID NO: 111 AAV CLv-R9 721 U.S. Pat. No. 8,734,809 SEQ ID NO:
112 AAV CLg-F1 722 U.S. Pat. No. 8,734,809 SEQ ID NO: 113 AAV
CLg-F2 723 U.S. Pat. No. 8,734,809 SEQ ID NO: 114 AAV CLg-F3 724
U.S. Pat. No. 8,734,809 SEQ ID NO: 115 AAV CLg-F4 725 U.S. Pat. No.
8,734,809 SEQ ID NO: 116 AAV CLg-F5 726 U.S. Pat. No. 8,734,809 SEQ
ID NO: 117 AAV CLg-F6 727 U.S. Pat. No. 8,734,809 SEQ ID NO: 117
AAV CLg-F7 728 U.S. Pat. No. 8,734,809 SEQ ID NO: 118 AAV CLg-F8
729 U.S. Pat. No. 8,734,809 SEQ ID NO: 117 AAV CSp-1 730 U.S. Pat.
No. 8,734,809 SEQ ID NO: 119 AAV CSp-10 731 U.S. Pat. No. 8,734,809
SEQ ID NO: 120 AAV CSp-11 732 U.S. Pat. No. 8,734,809 SEQ ID NO:
121 AAV CSp-2 733 U.S. Pat. No. 8,734,809 SEQ ID NO: 122 AAV CSp-3
734 U.S. Pat. No. 8,734,809 SEQ ID NO: 123 AAV CSp-4 735 U.S. Pat.
No. 8,734,809 SEQ ID NO: 124 AAV CSp-6 736 U.S. Pat. No. 8,734,809
SEQ ID NO: 125 AAV CSp-7 737 U.S. Pat. No. 8,734,809 SEQ ID NO: 126
AAV CSp-8 738 U.S. Pat. No. 8,734,809 SEQ ID NO: 127 AAV CSp-9 739
U.S. Pat. No. 8,734,809 SEQ ID NO: 128 AAV CHt-2 740 U.S. Pat. No.
8,734,809 SEQ ID NO: 129 AAV CHt-3 741 U.S. Pat. No. 8,734,809 SEQ
ID NO: 130 AAV CKd-1 742 U.S. Pat. No. 8,734,809 SEQ ID NO: 131 AAV
CKd-10 743 U.S. Pat. No. 8,734,809 SEQ ID NO: 132 AAV CKd-2 744
U.S. Pat. No. 8,734,809 SEQ ID NO: 133 AAV CKd-3 745 U.S. Pat. No.
8,734,809 SEQ ID NO: 134 AAV CKd-4 746 U.S. Pat. No. 8,734,809 SEQ
ID NO: 135 AAV CKd-6 747 U.S. Pat. No. 8,734,809 SEQ ID NO: 136 AAV
CKd-7 748 U.S. Pat. No. 8,734,809 SEQ ID NO: 137 AAV CKd-8 749 U.S.
Pat. No. 8,734,809 SEQ ID NO: 138 AAV CLv-1 750 U.S. Pat. No.
8,734,809 SEQ ID NO: 139 AAV CLv-12 751 U.S. Pat. No. 8,734,809 SEQ
ID NO: 140 AAV CLv-13 752 U.S. Pat. No. 8,734,809 SEQ ID NO: 141
AAV CLv-2 753 U.S. Pat. No. 8,734,809 SEQ ID NO: 142 AAV CLv-3 754
U.S. Pat. No. 8,734,809 SEQ ID NO: 143 AAV CLv-4 755 U.S. Pat. No.
8,734,809 SEQ ID NO: 144 AAV CLv-6 756 U.S. Pat. No. 8,734,809 SEQ
ID NO: 145 AAV CLv-8 757 U.S. Pat. No. 8,734,809 SEQ ID NO: 146 AAV
CKd-B1 758 U.S. Pat. No. 8,734,809 SEQ ID NO: 147 AAV CKd-B2 759
U.S. Pat. No. 8,734,809 SEQ ID NO: 148 AAV CKd-B3 760 U.S. Pat. No.
8,734,809 S EQ ID NO: 149 AAV CKd-B4 761 U.S. Pat. No. 8,734,809
SEQ ID NO: 150 AAV CKd-B5 762 U.S. Pat. No. 8,734,809 SEQ ID NO:
151 AAV CKd-B6 763 U.S. Pat. No. 8,734,809 SEQ ID NO: 152 AAV
CKd-B7 764 U.S. Pat. No. 8,734,809 SEQ ID NO: 153 AAV CKd-B8 765
U.S. Pat. No. 8,734,809 SEQ ID NO: 154 AAV CKd-H1 766 U.S. Pat. No.
8,734,809 SEQ ID NO: 155 AAV CKd-H2 767 U.S. Pat. No. 8,734,809 SEQ
ID NO: 156 AAV CKd-H3 768 U.S. Pat. No. 8,734,809 SEQ ID NO: 157
AAV CKd-H4 769 U.S. Pat. No. 8,734,809 SEQ ID NO: 158 AAV CKd-H5
770 U.S. Pat. No. 8,734,809 SEQ ID NO: 159 AAV CKd-H6 771 U.S. Pat.
No. 8,734,809 S EQ ID NO: 151 AAV CHt-1 772 U.S. Pat. No. 8,734,809
SEQ ID NO: 160 AAV CHt-P2 773 WO2016065001 SEQ ID NO: 1 AAV CHt-P5
774 WO2016065001 SEQ ID NO: 2 AAV CHt-P9 775 WO2016065001 SEQ ID
NO: 3 AAV CBr-7.1 776 WO2016065001 SEQ ID NO: 4 AAV CBr-7.2 777
WO2016065001 SEQ ID NO: 5 AAV CBr-7.3 778 WO2016065001 SEQ ID NO: 6
AAV CBr-7.4 779 WO2016065001 SEQ ID NO: 7 AAV CBr-7.5 780
WO2016065001 SEQ ID NO: 8 AAV CBr-7.7 781 WO2016065001 SEQ ID NO: 9
AAV CBr-7.8 782 WO2016065001 SEQ ID NO: 10 AAV CBr-7.10 783
WO2016065001 SEQ ID NO: 11 AAV CKd-N3 784 WO2016065001 SEQ ID NO:
12 AAV CKd-N4 785 WO2016065001 SEQ ID NO: 13 AAV CKd-N9 786
WO2016065001 SEQ ID NO: 14 AAV CLv-L4 787 WO2016065001 SEQ ID NO:
15 AAV CLv-L5 788 WO2016065001 SEQ ID NO: 16 AAV CLv-L6 789
WO2016065001 SEQ ID NO: 17 AAV CLv-K1 790 WO2016065001 SEQ ID NO:
18 AAV CLv-K3 791 WO2016065001 SEQ ID NO: 19 AAV CLv-K6 792
WO2016065001 SEQ ID NO: 20 AAV CLv-M1 793 WO2016065001 SEQ ID NO:
21 AAV CLv-M11 794 WO2016065001 SEQ ID NO: 22 AAV CLv-M2 795
WO2016065001 SEQ ID NO: 23 AAV CLv-M5 796 WO2016065001 SEQ ID NO:
24 AAV CLv-M6 797 WO2016065001 SEQ ID NO: 25 AAV CLv-M7 798
WO2016065001 SEQ ID NO: 26 AAV CLv-M8 799 WO2016065001 SEQ ID NO:
27 AAV CLv-M9 800 WO2016065001 SEQ ID NO: 28 AAV CHt-P1 801
WO2016065001 SEQ ID NO: 29 AAV CHt-P6 802 WO2016065001 SEQ ID NO:
30 AAV CHt-P8 803 WO2016065001 SEQ ID NO: 31 AAV CHt-6.1 804
WO2016065001 SEQ ID NO: 32 AAV CHt-6.10 805 WO2016065001 SEQ ID NO:
33 AAV CHt-6.5 806 WO2016065001 SEQ ID NO: 34 AAV CHt-6.6 807
WO2016065001 SEQ ID NO: 35 AAV CHt-6.7 808 WO2016065001 SEQ ID NO:
36 AAV CHt-6.8 809 WO2016065001 SEQ ID NO: 37 AAV CSp-8.10 810
WO2016065001 SEQ ID NO: 38 AAV CSp-8.2 811 WO2016065001 SEQ ID NO:
39 AAV CSp-8.4 812 WO2016065001 SEQ ID NO: 40 AAV CSp-8.5 813
WO2016065001 SEQ ID NO: 41 AAV CSp-8.6 814 WO2016065001 SEQ ID NO:
42 AAV CSp-8.7 815 WO2016065001 SEQ ID NO: 43 AAV CSp-8.8 816
WO2016065001 SEQ ID NO: 44 AAV CSp-8.9 817 WO2016065001 SEQ ID NO:
45 AAV CBr-B7.3 818 WO2016065001 SEQ ID NO: 46 AAV CBr-B7.4 819
WO2016065001 SEQ ID NO: 47 AAV3B 820 WO2016065001 SEQ ID NO: 48
AAV4 821 WO2016065001 SEQ ID NO: 49 AAV5 822 WO2016065001 SEQ ID
NO: 50 AAV CHt-P2 823 WO2016065001 SEQ ID NO: 51 AAV CHt-P5 824
WO2016065001 SEQ ID NO: 52 AAV CHt-P9 825 WO2016065001 SEQ ID NO:
53 AAV CBr-7.1 826 WO2016065001 SEQ ID NO: 54 AAV CBr-7.2 827
WO2016065001 SEQ ID NO: 55 AAV CBr-7.3 828 WO2016065001 SEQ ID NO:
56 AAV CBr-7.4 829 WO2016065001 SEQ ID NO: 57 AAV CBr-7.5 830
WO2016065001 SEQ ID NO: 58 AAV CBr-7.7 831 WO2016065001 SEQ ID NO:
59 AAV CBr-7.8 832 WO2016065001 SEQ ID NO: 60 AAV CBr-7.10 833
WO2016065001 SEQ ID NO: 61 AAV CKd-N3 834 WO2016065001 SEQ ID NO:
62 AAV CKd-N4 835 WO2016065001 SEQ ID NO: 63 AAV CKd-N9 836
WO2016065001 SEQ ID NO: 64 AAV CLv-L4 837 WO2016065001 SEQ ID NO:
65 AAV CLv-L5 838 WO2016065001 SEQ ID NO: 66 AAV CLv-L6 839
WO2016065001 SEQ ID NO: 67 AAV CLv-K1 840 WO2016065001 SEQ ID NO:
68 AAV CLv-K3 841 WO2016065001 SEQ ID NO: 69 AAV CLv-K6 842
WO2016065001 SEQ ID NO: 70 AAV CLv-M1 843 WO2016065001 SEQ ID NO:
71 AAV CLv-M11 844 WO2016065001 SEQ ID NO: 72 AAV CLv-M2 845
WO2016065001 SEQ ID NO: 73 AAV CLv-M5 846 WO2016065001 SEQ ID NO:
74 AAV CLv-M6 847 WO2016065001 SEQ ID NO: 75 AAV CLv-M7 848
WO2016065001 SEQ ID NO: 76 AAV CLv-M8 849 WO2016065001 SEQ ID NO:
77 AAV CLv-M9 850 WO2016065001 SEQ ID NO: 78 AAV CHt-P1 851
WO2016065001 SEQ ID NO: 79 AAV CHt-P6 852 WO2016065001 SEQ ID NO:
80 AAV CHt-P8 853 WO2016065001 SEQ ID NO: 81 AAV CHt-6.1 854
WO2016065001 SEQ ID NO: 82 AAV CHt-6.10 855 WO2016065001 SEQ ID NO:
83 AAV CHt-6.5 856 WO2016065001 SEQ ID NO: 84 AAV CHt-6.6 857
WO2016065001 SEQ ID NO: 85 AAV CHt-6.7 858 WO2016065001 SEQ ID NO:
86 AAV CHt-6.8 859 WO2016065001 SEQ ID NO: 87 AAV CSp-8.10 860
WO2016065001 SEQ ID NO: 88 AAV CSp-8.2 861 WO2016065001 SEQ ID NO:
89 AAV CSp-8.4 862 WO2016065001 SEQ ID NO: 90 AAV CSp-8.5 863
WO2016065001 SEQ ID NO: 91 AAV CSp-8.6 864 WO2016065001 SEQ ID NO:
92 AAV CSp-8.7 865 WO2016065001 SEQ ID NO: 93 AAV CSp-8.8 866
WO2016065001 SEQ ID NO: 94 AAV CSp-8.9 867 WO2016065001 SEQ ID NO:
95 AAV CBr-B7.3 868 WO2016065001 SEQ ID NO: 96
AAV CBr-B7.4 869 WO2016065001 SEQ ID NO: 97 AAV3B 870 WO2016065001
SEQ ID NO: 98 AAV4 871 WO2016065001 SEQ ID NO: 99 AAV5 872
WO2016065001 SEQ ID NO: 100 PHP.N/PHP.B-DGT 873 WO2017100671 SEQ ID
NO: 46 PHP.S/G2A12 874 WO2017100671 SEQ ID NO: 47 AAV9/hu.14 K449R
875 WO2017100671 SEQ ID NO: 45 GPV 992 US9624274B2 SEQ ID NO: 192
B19 993 US9624274B2 SEQ ID NO: 193 MVM 994 US9624274B2 SEQ ID NO:
194 FPV 995 US9624274B2 SEQ ID NO: 195 CPV 996 US9624274B2 SEQ ID
NO: 196 AAV6 997 US9546112B2 SEQ ID NO: 5 AAV6 998 US9457103B2 SEQ
ID NO: 1 AAV2 999 US9457103B2 SEQ ID NO: 2 ShH10 1000 US9457103B2
SEQ ID NO: 3 ShH13 1001 US9457103B2 SEQ ID NO: 4 ShH10 1002
US9457103B2 SEQ ID NO: 5 ShH10 1003 US9457103B2 SEQ ID NO: 6 ShH10
1004 US9457103B2 SEQ ID NO: 7 ShH10 1005 US9457103B2 SEQ ID NO: 8
ShH10 1006 US9457103B2 SEQ ID NO: 9 rh74 1007 US9434928B2 SEQ ID
NO: 1, US2015023924A1 SEQ ID NO: 2 rh74 1008 US9434928B2 SEQ ID NO:
2, US2015023924A1 SEQ ID NO: 1 AAV8 1009 US9434928B2 SEQ ID NO: 4
rh74 1010 US9434928B2 SEQ ID NO: 5 rh74 (RHM4-1) 1011
US2015023924A1 SEQ ID NO: 5, US20160375110A1 SEQ ID NO: 4 rh74
(RHM15-1) 1012 US2015023924A1 SEQ ID NO: 6, US20160375110A1 SEQ ID
NO: 5 rh74 (RHM15-2) 1013 US2015023924A1 SEQ ID NO: 7,
US20160375110A1 SEQ ID NO: 6 rh74 (RHM15-3/ 1014 US2015023924A1 SEQ
ID NO: 8, RHM15-5) US20160375110A1 SEQ ID NO: 7 rh74 (RHM15-4) 1015
US2015023924A1 SEQ ID NO: 9, US20160375110A1 SEQ ID NO: 8 rh74
(RHM15-6) 1016 US2015023924A1 SEQ ID NO: 10, US20160375110A1 SEQ ID
NO: 9 rh74 (RHM4-1) 1017 US2015023924A1 SEQ ID NO: 11 rh74
(RHM15-1) 1018 US2015023924A1 SEQ ID NO: 12 rh74 (RHM15-2) 1019
US2015023924A1 SEQ ID NO: 13 rh74 (RHM15-3/ 1020 US2015023924A1 SEQ
ID NO: 14 RHM15-5) rh74 (RHM15-4) 1021 US2015023924A1 SEQ ID NO: 15
rh74 (RHM15-6) 1022 US2015023924A1 SEQ ID NO: 16 AAV2 (comprising
lung 1023 US20160175389A1 SEQ ID NO: 9 specific polypeptide) AAV2
(comprising lung 1024 US20160175389A1 SEQ ID NO: 10 specific
polypeptide) Anc80 1025 US20170051257A1 SEQ ID NO: 1 Anc80 1026
US20170051257A1 SEQ ID NO: 2 Anc81 1027 US20170051257A1 SEQ ID NO:
3 Anc80 1028 US20170051257A1 SEQ ID NO: 4 Anc82 1029
US20170051257A1 SEQ ID NO: 5 Anc82 1030 US20170051257A1 SEQ ID NO:
6 Anc83 1031 US20170051257A1 SEQ ID NO: 7 Anc83 1032
US20170051257A1 SEQ ID NO: 8 Anc84 1033 US20170051257A1 SEQ ID NO:
9 Anc84 1034 US20170051257A1 SEQ ID NO: 10 Anc94 1035
US20170051257A1 SEQ ID NO: 11 Anc94 1036 US20170051257A1 SEQ ID NO:
12 Anc113 1037 US20170051257A1 SEQ ID NO: 13 Anc113 1038
US20170051257A1 SEQ ID NO: 14 Anc126 1039 US20170051257A1 SEQ ID
NO: 15 Anc126 1040 US20170051257A1 SEQ ID NO: 16 Anc127 1041
US20170051257A1 SEQ ID NO: 17 Anc127 1042 US20170051257A1 SEQ ID
NO: 18 Anc80L27 1043 US20170051257A1 SEQ ID NO: 19 Anc80L59 1044
US20170051257A1 SEQ ID NO: 20 Anc80L60 1045 US20170051257A1 SEQ ID
NO: 21 Anc80L62 1046 US20170051257A1 SEQ ID NO: 22 Anc80L65 1047
US20170051257A1 SEQ ID NO: 23 Anc80L33 1048 US20170051257A1 SEQ ID
NO: 24 Anc80L36 1049 US20170051257A1 SEQ ID NO: 25 Anc80L44 1050
US20170051257A1 SEQ ID NO: 26 Anc80L1 1051 US20170051257A1 SEQ ID
NO: 35 Anc80L1 1052 US20170051257A1 SEQ ID NO: 36 AAV-X1 1053
US8283151B2 SEQ ID NO: 11 AAV-X1b 1054 US8283151B2 SEQ ID NO: 12
AAV-X5 1055 US8283151B2 SEQ ID NO: 13 AAV-X19 1056 US8283151B2 SEQ
ID NO: 14 AAV-X21 1057 US8283151B2 SEQ ID NO: 15 AAV-X22 1058
US8283151B2 SEQ ID NO: 16 AAV-X23 1059 US8283151B2 SEQ ID NO: 17
AAV-X24 1060 US8283151B2 SEQ ID NO: 18 AAV-X25 1061 US8283151B2 SEQ
ID NO: 19 AAV-X26 1062 US8283151B2 SEQ ID NO: 20 AAV-X1 1063
US8283151B2 SEQ ID NO: 21 AAV-X1b 1064 US8283151B2 SEQ ID NO: 22
AAV-X5 1065 US8283151B2 SEQ ID NO: 23 AAV-X19 1066 US8283151B2 SEQ
ID NO: 24 AAV-X21 1067 US8283151B2 SEQ ID NO: 25 AAV-X22 1068
US8283151B2 SEQ ID NO: 26 AAV-X23 1069 US8283151B2 SEQ ID NO: 27
AAV-X24 1070 US8283151B2 SEQ ID NO: 28 AAV-X25 1071 US8283151B2 SEQ
ID NO: 29 AAV-X26 1072 US8283151B2 SEQ ID NO: 30 AAVrh8 1073
WO2016054554A1 SEQ ID NO: 8 AAVrh8VP2FC5 1074 WO2016054554A1 SEQ ID
NO: 9 AAVrh8VP2FC44 1075 WO2016054554A1 SEQ ID NO: 10
AAVrh8VP2ApoB100 1076 WO2016054554A1 SEQ ID NO: 11 AAVrh8VP2RVG
1077 WO2016054554A1 SEQ ID NO: 12 AAVrh8VP2Angiopep-2 1078
WO2016054554A1 SEQ ID NO: 13 VP2 AAV9.47VP1.3 1079 WO2016054554A1
SEQ ID NO: 14 AAV9.47VP2ICAMg3 1080 WO2016054554A1 SEQ ID NO: 15
AAV9.47VP2RVG 1081 WO2016054554A1 SEQ ID NO: 16 AAV9.47VP2Angiopep-
1082 WO2016054554A1 SEQ ID NO: 17 2 AAV9.47VP2A-string 1083
WO2016054554A1 SEQ ID NO: 18 AAVrh8VP2FC5 VP2 1084 WO2016054554A1
SEQ ID NO: 19 AAVrh8VP2FC44 VP2 1085 WO2016054554A1 SEQ ID NO: 20
AAVrh8VP2ApoB100 1086 WO2016054554A1 SEQ ID NO: 21 VP2 AAVrh8VP2RVG
VP2 1087 WO2016054554A1 SEQ ID NO: 22 AAVrh8VP2Angiopep-2 1088
WO2016054554A1 SEQ ID NO: 23 VP2 AAV9.47VP2ICAMg3 1089
WO2016054554A1 SEQ ID NO: 24 VP2 AAV9.47VP2RVG VP2 1090
WO2016054554A1 SEQ ID NO: 25 AAV9.47VP2Angiopep-2 1091
WO2016054554A1 SEQ ID NO: 26 VP2 AAV9.47VP2A-string 1092
WO2016054554A1 SEQ ID NO: 27 VP2 rAAV-B1 1093 WO2016054557A1 SEQ ID
NO: 1 rAAV-B2 1094 WO2016054557A1 SEQ ID NO: 2 rAAV-B3 1095
WO2016054557A1 SEQ ID NO: 3 rAAV-B4 1096 WO2016054557A1 SEQ ID NO:
4 rAAV-B1 1097 WO2016054557A1 SEQ ID NO: 5 rAAV-B2 1098
WO2016054557A1 SEQ ID NO: 6 rAAV-B3 1099 WO2016054557A1 SEQ ID NO:
7 rAAV-B4 1100 WO2016054557A1 SEQ ID NO: 8 rAAV-L1 1101
WO2016054557A1 SEQ ID NO: 9 rAAV-L2 1102 WO2016054557A1 SEQ ID NO:
10 rAAV-L3 1103 WO2016054557A1 SEQ ID NO: 11 rAAV-L4 1104
WO2016054557A1 SEQ ID NO: 12 rAAV-L1 1105 WO2016054557A1 SEQ ID NO:
13 rAAV-L2 1106 WO2016054557A1 SEQ ID NO: 14 rAAV-L3 1107
WO2016054557A1 SEQ ID NO: 15 rAAV-L4 1108 WO2016054557A1 SEQ ID NO:
16 AAV9 1109 WO2016073739A1 SEQ ID NO: 3 rAAV 1110 WO2016081811A1
SEQ ID NO: 1 rAAV 1111 WO2016081811A1 SEQ ID NO: 2 rAAV 1112
WO2016081811A1 SEQ ID NO: 3 rAAV 1113 WO2016081811A1 SEQ ID NO: 4
rAAV 1114 WO2016081811A1 SEQ ID NO: 5 rAAV 1115 WO2016081811A1 SEQ
ID NO: 6 rAAV 1116 WO2016081811A1 SEQ ID NO: 7 rAAV 1117
WO2016081811A1 SEQ ID NO: 8 rAAV 1118 WO2016081811A1 SEQ ID NO: 9
rAAV 1119 WO2016081811A1 SEQ ID NO: 10 rAAV 1120 WO2016081811A1 SEQ
ID NO: 11 rAAV 1121 WO2016081811A1 SEQ ID NO: 12 rAAV 1122
WO2016081811A1 SEQ ID NO: 13 rAAV 1123 WO2016081811A1 SEQ ID NO: 14
rAAV 1124 WO2016081811A1 SEQ ID NO: 15 rAAV 1125 WO2016081811A1 SEQ
ID NO: 16 rAAV 1126 WO2016081811A1 SEQ ID NO: 17 rAAV 1127
WO2016081811A1 SEQ ID NO: 18 rAAV 1128 WO2016081811A1 SEQ ID NO: 19
rAAV 1129 WO2016081811A1 SEQ ID NO: 20 rAAV 1130 WO2016081811A1 SEQ
ID NO: 21 rAAV 1131 WO2016081811A1 SEQ ID NO: 22 rAAV 1132
WO2016081811A1 SEQ ID NO: 23 rAAV 1133 WO2016081811A1 SEQ ID NO: 24
rAAV 1134 WO2016081811A1 SEQ ID NO: 25 rAAV 1135 WO2016081811A1 SEQ
ID NO: 26 rAAV 1136 WO2016081811A1 SEQ ID NO: 27 rAAV 1137
WO2016081811A1 SEQ ID NO: 28 rAAV 1138 WO2016081811A1 SEQ ID NO: 29
rAAV 1139 WO2016081811A1 SEQ ID NO: 30 rAAV 1140 WO2016081811A1 SEQ
ID NO: 31 rAAV 1141 WO2016081811A1 SEQ ID NO: 32 rAAV 1142
WO2016081811A1 SEQ ID NO: 33 rAAV 1143 WO2016081811A1 SEQ ID NO: 34
rAAV 1144 WO2016081811A1 SEQ ID NO: 35 rAAV 1145 WO2016081811A1 SEQ
ID NO: 36 rAAV 1146 WO2016081811A1 SEQ ID NO: 37 rAAV 1147
WO2016081811A1 SEQ ID NO: 38 rAAV 1148 WO2016081811A1 SEQ ID NO: 39
rAAV 1149 WO2016081811A1 SEQ ID NO: 40 rAAV 1150 WO2016081811A1 SEQ
ID NO: 41 rAAV 1151 WO2016081811A1 SEQ ID NO: 42 rAAV 1152
WO2016081811A1 SEQ ID NO: 43 rAAV 1153 WO2016081811A1 SEQ ID NO: 44
rAAV 1154 WO2016081811A1 SEQ ID NO: 45 rAAV 1155 WO2016081811A1 SEQ
ID NO: 46 rAAV 1156 WO2016081811A1 SEQ ID NO: 47 rAAV 1157
WO2016081811A1 SEQ ID NO: 48 rAAV 1158 WO2016081811A1 SEQ ID NO: 49
rAAV 1159 WO2016081811A1 SEQ ID NO: 50 rAAV 1160 WO2016081811A1 SEQ
ID NO: 51 rAAV 1161 WO2016081811A1 SEQ ID NO: 52 rAAV 1162
WO2016081811A1 SEQ ID NO: 53 rAAV 1163 WO2016081811A1 SEQ ID NO: 54
rAAV 1164 WO2016081811A1 SEQ ID NO: 55 rAAV 1165 WO2016081811A1 SEQ
ID NO: 56 rAAV 1166 WO2016081811A1 SEQ ID NO: 57 rAAV 1167
WO2016081811A1 SEQ ID NO: 58 rAAV 1168 WO2016081811A1 SEQ ID NO: 59
rAAV 1169 WO2016081811A1 SEQ ID NO: 60 rAAV 1170 WO2016081811A1 SEQ
ID NO: 61 rAAV 1171 WO2016081811A1 SEQ ID NO: 62 rAAV 1172
WO2016081811A1 SEQ ID NO: 63 rAAV 1173 WO2016081811A1 SEQ ID NO: 64
rAAV 1174 WO2016081811A1 SEQ ID NO: 65 rAAV 1175 WO2016081811A1 SEQ
ID NO: 66 rAAV 1176 WO2016081811A1 SEQ ID NO: 67 rAAV 1177
WO2016081811A1 SEQ ID NO: 68 rAAV 1178 WO2016081811A1 SEQ ID NO: 69
rAAV 1179 WO2016081811A1 SEQ ID NO: 70 rAAV 1180 WO2016081811A1 SEQ
ID NO: 71 rAAV 1181 WO2016081811A1 SEQ ID NO: 72 rAAV 1182
WO2016081811A1 SEQ ID NO: 73 rAAV 1183 WO2016081811A1 SEQ ID NO: 74
rAAV 1184 WO2016081811A1 SEQ ID NO: 75 rAAV 1185 WO2016081811A1 SEQ
ID NO: 76 rAAV 1186 WO2016081811A1 SEQ ID NO: 77 rAAV 1187
WO2016081811A1 SEQ ID NO: 78 rAAV 1188 WO2016081811A1 SEQ ID NO: 79
rAAV 1189 WO2016081811A1 SEQ ID NO: 80 rAAV 1190 WO2016081811A1 SEQ
ID NO: 81 rAAV 1191 WO2016081811A1 SEQ ID NO: 82 rAAV 1192
WO2016081811A1 SEQ ID NO: 83 rAAV 1193 WO2016081811A1 SEQ ID NO: 84
rAAV 1194 WO2016081811A1 SEQ ID NO: 85 rAAV 1195 WO2016081811A1 SEQ
ID NO: 86 rAAV 1196 WO2016081811A1 SEQ ID NO: 87 rAAV 1197
WO2016081811A1 SEQ ID NO: 88 rAAV 1198 WO2016081811A1 SEQ ID NO: 89
rAAV 1199 WO2016081811A1 SEQ ID NO: 90 rAAV 1200 WO2016081811A1 SEQ
ID NO: 91 rAAV 1201 WO2016081811A1 SEQ ID NO: 92 rAAV 1202
WO2016081811A1 SEQ ID NO: 93 rAAV 1203 WO2016081811A1 SEQ ID NO: 94
rAAV 1204 WO2016081811A1 SEQ ID NO: 95 rAAV 1205 WO2016081811A1 SEQ
ID NO: 96 rAAV 1206 WO2016081811A1 SEQ ID NO: 97 rAAV 1207
WO2016081811A1 SEQ ID NO: 98 rAAV 1208 WO2016081811A1 SEQ ID NO: 99
rAAV 1209 WO2016081811A1 SEQ ID NO: 100 rAAV 1210 WO2016081811A1
SEQ ID NO: 101 rAAV 1211 WO2016081811A1 SEQ ID NO: 102 rAAV 1212
WO2016081811A1 SEQ ID NO: 103 rAAV 1213 WO2016081811A1 SEQ ID NO:
104 rAAV 1214 WO2016081811A1 SEQ ID NO: 105 rAAV 1215
WO2016081811A1 SEQ ID NO: 106 rAAV 1216 WO2016081811A1 SEQ ID NO:
107 rAAV 1217 WO2016081811A1 SEQ ID NO: 108
rAAV 1218 WO2016081811A1 SEQ ID NO: 109 rAAV 1219 WO2016081811A1
SEQ ID NO: 110 rAAV 1220 WO2016081811A1 SEQ ID NO: 111 rAAV 1221
WO2016081811A1 SEQ ID NO: 112 rAAV 1222 WO2016081811A1 SEQ ID NO:
113 rAAV 1223 WO2016081811A1 SEQ ID NO: 114 rAAV 1224
WO2016081811A1 SEQ ID NO: 115 rAAV 1225 WO2016081811A1 SEQ ID NO:
116 rAAV 1226 WO2016081811A1 SEQ ID NO: 117 rAAV 1227
WO2016081811A1 SEQ ID NO: 118 rAAV 1228 WO2016081811A1 SEQ ID NO:
119 rAAV 1229 WO2016081811A1 SEQ ID NO: 120 rAAV 1230
WO2016081811A1 SEQ ID NO: 121 rAAV 1231 WO2016081811A1 SEQ ID NO:
122 rAAV 1232 WO2016081811A1 SEQ ID NO: 123 rAAV 1233
WO2016081811A1 SEQ ID NO: 124 rAAV 1234 WO2016081811A1 SEQ ID NO:
125 rAAV 1235 WO2016081811A1 SEQ ID NO: 126 rAAV 1236
WO2016081811A1 SEQ ID NO: 127 rAAV 1237 WO2016081811A1 SEQ ID NO:
128 AAV8 E532K 1238 WO2016081811A1 SEQ ID NO: 133 AAV8 E532K 1239
WO2016081811A1 SEQ ID NO: 134 rAAV4 1240 WO2016115382A1 SEQ ID NO:
2 rAAV4 1241 WO2016115382A1 SEQ ID NO: 3 rAAV4 1242 WO2016115382A1
SEQ ID NO: 4 rAAV4 1243 WO2016115382A1 SEQ ID NO: 5 rAAV4 1244
WO2016115382A1 SEQ ID NO: 6 rAAV4 1245 WO2016115382A1 SEQ ID NO: 7
rAAV4 1246 WO2016115382A1 SEQ ID NO: 8 rAAV4 1247 WO2016115382A1
SEQ ID NO: 9 rAAV4 1248 WO2016115382A1 SEQ ID NO: 10 rAAV4 1249
WO2016115382A1 SEQ ID NO: 11 rAAV4 1250 WO2016115382A1 SEQ ID NO:
12 rAAV4 1251 WO2016115382A1 SEQ ID NO: 13 rAAV4 1252
WO2016115382A1 SEQ ID NO: 14 rAAV4 1253 WO2016115382A1 SEQ ID NO:
15 rAAV4 1254 WO2016115382A1 SEQ ID NO: 16 rAAV4 1255
WO2016115382A1 SEQ ID NO: 17 rAAV4 1256 WO2016115382A1 SEQ ID NO:
18 rAAV4 1257 WO2016115382A1 SEQ ID NO: 19 rAAV4 1258
WO2016115382A1 SEQ ID NO: 20 rAAV4 1259 WO2016115382A1 SEQ ID NO:
21 AAV11 1260 WO2016115382A1 SEQ ID NO: 22 AAV12 1261
WO2016115382A1 SEQ ID NO: 23 rh32 1262 WO2016115382A1 SEQ ID NO: 25
rh33 1263 WO2016115382A1 SEQ ID NO: 26 rh34 1264 WO2016115382A1 SEQ
ID NO: 27 rAAV4 1265 WO2016115382A1 SEQ ID NO: 28 rAAV4 1266
WO2016115382A1 SEQ ID NO: 29 rAAV4 1267 WO2016115382A1 SEQ ID NO:
30 rAAV4 1268 WO2016115382A1 SEQ ID NO: 31 rAAV4 1269
WO2016115382A1 SEQ ID NO: 32 rAAV4 1270 WO2016115382A1 SEQ ID NO:
33 AAV2/8 1271 WO2016131981A1 SEQ ID NO: 47 AAV2/8 1272
WO2016131981A1 SEQ ID NO: 48 ancestral AAV 1273 WO2016154344A1 SEQ
ID NO: 7 ancestral AAV variant 1274 WO2016154344A1 SEQ ID NO: 13 C4
ancestral AAV variant 1275 WO2016154344A1 SEQ ID NO: 14 C7
ancestral AAV variant 1276 WO2016154344A1 SEQ ID NO: 15 G4
consensus amino acid 1277 WO2016154344A1 SEQ ID NO: 16 sequence of
ancestral AAV variants, C4, C7 and G4 consensus amino acid 1278
WO2016154344A1 SEQ ID NO: 17 sequence of ancestral AAV variants, C4
and C7 AAV8 (with a AAV2 1279 WO2016150403A1 SEQ ID NO: 13
phospholipase domain) AAV VR-942n 1280 US20160289275A1 SEQ ID NO:
10 AAV5-A (M569V) 1281 US20160289275A1 SEQ ID NO: 13 AAV5-A (M569V)
1282 US20160289275A1 SEQ ID NO: 14 AAV5-A (Y585V) 1283
US20160289275A1 SEQ ID NO: 16 AAV5-A (Y585V) 1284 US20160289275A1
SEQ ID NO: 17 AAV5-A (L587T) 1285 US20160289275A1 SEQ ID NO: 19
AAV5-A (L587T) 1286 US20160289275A1 SEQ ID NO: 20 AAV5-A (Y585V/
1287 US20160289275A1 SEQ ID NO: 22 L587T) AAV5-A (Y585V/ 1288
US20160289275A1 SEQ ID NO: 23 L587T) AAV5-B (D652A) 1289
US20160289275A1 SEQ ID NO: 25 AAV5-B (D652A) 1290 US20160289275A1
SEQ ID NO: 26 AAV5-B (T362M) 1291 US20160289275A1 SEQ ID NO: 28
AAV5-B (T362M) 1292 US20160289275A1 SEQ ID NO: 29 AAV5-B (Q359D)
1293 US20160289275A1 SEQ ID NO: 31 AAV5-B (Q359D) 1294
US20160289275A1 SEQ ID NO: 32 AAV5-B (E350Q) 1295 US20160289275A1
SEQ ID NO: 34 AAV5-B (E350Q) 1296 US20160289275A1 SEQ ID NO: 35
AAV5-B (P533S) 1297 US20160289275A1 SEQ ID NO: 37 AAV5-B (P533S)
1298 US20160289275A1 SEQ ID NO: 38 AAV5-B (P533G) 1299
US20160289275A1 SEQ ID NO: 40 AAV5-B (P533G) 1300 US20160289275A1
SEQ ID NO: 41 AAV5-mutation in 1301 US20160289275A1 SEQ ID NO: 43
loop VII AAV5-mutation in 1302 US20160289275A1 SEQ ID NO: 44 loop
VII AAV8 1303 US20160289275A1 SEQ ID NO: 47 Mut A 1304
WO2016181123A1 SEQ ID NO: 1 (LK03/AAV8) Mut B 1305 WO2016181123A1
SEQ ID NO: 2 (LK03/AAV5) Mut C 1306 WO2016181123A1 SEQ ID NO: 3
(AAV8/AAV3B) Mut D 1307 WO2016181123A1 SEQ ID NO: 4 (AAV5/AAV3B)
Mut E 1308 WO2016181123A1 SEQ ID NO: 5 (AAV8/AAV3B) Mut F 1309
WO2016181123A1 SEQ ID NO: 6 (AAV3B/AAV8) AAV44.9 1310
WO2016183297A1 SEQ ID NO: 4 AAV44.9 1311 WO2016183297A1 SEQ ID NO:
5 AAVrh8 1312 WO2016183297A1 SEQ ID NO: 6 AAV44.9 (S470N) 1313
WO2016183297A1 SEQ ID NO: 9 rh74 VP1 1314 US20160375110A1 SEQ ID
NO: 1 AAV-LK03 (L125I) 1315 WO2017015102A1 SEQ ID NO: 5 AAV3B
(S663V + 1316 WO2017015102A1 SEQ ID NO: 6 T492V) Anc80 1317
WO2017019994A2 SEQ ID NO: 1 Anc80 1318 WO2017019994A2 SEQ ID NO: 2
Anc81 1319 WO2017019994A2 SEQ ID NO: 3 Anc81 1320 WO2017019994A2
SEQ ID NO: 4 Anc82 1321 WO2017019994A2 SEQ ID NO: 5 Anc82 1322
WO2017019994A2 SEQ ID NO: 6 Anc83 1323 WO2017019994A2 SEQ ID NO: 7
Anc83 1324 WO2017019994A2 SEQ ID NO: 8 Anc84 1325 WO2017019994A2
SEQ ID NO: 9 Anc84 1326 WO2017019994A2 SEQ ID NO: 10 Anc94 1327
WO2017019994A2 SEQ ID NO: 11 Anc94 1328 WO2017019994A2 SEQ ID NO:
12 Anc113 1329 WO2017019994A2 SEQ ID NO: 13 Anc113 1330
WO2017019994A2 SEQ ID NO: 14 Anc126 1331 WO2017019994A2 SEQ ID NO:
15 Anc126 1332 WO2017019994A2 SEQ ID NO: 16 Anc127 1333
WO2017019994A2 SEQ ID NO: 17 Anc127 1334 WO2017019994A2 SEQ ID NO:
18 Anc80L27 1335 WO2017019994A2 SEQ ID NO: 19 Anc80L59 1336
WO2017019994A2 SEQ ID NO: 20 Anc80L60 1337 WO2017019994A2 SEQ ID
NO: 21 Anc80L62 1338 WO2017019994A2 SEQ ID NO: 22 Anc80L65 1339
WO2017019994A2 SEQ ID NO: 23 Anc80L33 1340 WO2017019994A2 SEQ ID
NO: 24 Anc80L36 1341 WO2017019994A2 SEQ ID NO: 25 Anc80L44 1342
WO2017019994A2 SEQ ID NO: 26 Anc80L1 1343 WO2017019994A2 SEQ ID NO:
35 Anc80L1 1344 WO2017019994A2 SEQ ID NO: 36 AAVrh10 1345
WO2017019994A2 SEQ ID NO: 41 Anc110 1346 WO2017019994A2 SEQ ID NO:
42 Anc110 1347 WO2017019994A2 SEQ ID NO: 43 AAVrh32.33 1348
WO2017019994A2 SEQ ID NO: 45 AAVrh74 1349 WO2017049031A1 SEQ ID NO:
1 AAV2 1350 WO2017053629A2 SEQ ID NO: 49 AAV2 1351 WO2017053629A2
SEQ ID NO: 50 AAV2 1352 WO2017053629A2 SEQ ID NO: 82 Parvo-like
virus 1353 WO2017070476A2 SEQ ID NO: 1 Parvo-like virus 1354
WO2017070476A2 SEQ ID NO: 2 Parvo-like virus 1355 WO2017070476A2
SEQ ID NO: 3 Parvo-like virus 1356 WO2017070476A2 SEQ ID NO: 4
Parvo-like virus 1357 WO2017070476A2 SEQ ID NO: 5 Parvo-like virus
1358 WO2017070476A2 SEQ ID NO: 6 AAVrh.10 1359 WO2017070516A1 SEQ
ID NO: 7 AAVrh.10 1360 WO2017070516A1 SEQ ID NO: 14 AAV2tYF 1361
WO2017070491A1 SEQ ID NO: 1 AAV-SPK 1362 WO2017075619A1 SEQ ID NO:
28 AAV2.5 1363 US20170128528A1 SEQ ID NO: 13 AAV1.1 1364
US20170128528A1 SEQ ID NO: 15 AAV6.1 1365 US20170128528A1 SEQ ID
NO: 17 AAV6.3.1 1366 US20170128528A1 SEQ ID NO: 18 AAV2i8 1367
US20170128528A1 SEQ ID NO: 28 AAV2i8 1368 US20170128528A1 SEQ ID
NO: 29 ttAAV 1369 US20170128528A1 SEQ ID NO: 30 ttAAV-S312N 1370
US20170128528A1 SEQ ID NO: 32 ttAAV-S312N 1371 US20170128528A1 SEQ
ID NO: 33 AAV6 (Y705, Y731, 1372 WO2016134337A1 SEQ ID NO: 24 and
T492) AAV2 1373 WO2016134375A1 SEQ ID NO: 9 AAV2 1374
WO2016134375A1 SEQ ID NO: 10
[0068] In one embodiment, the AAV serotype may be, or may have a
sequence as described in International Patent Publication
WO2015038958, the contents of which are herein incorporated by
reference in their entirety, such as, but not limited to, AAV9 (SEQ
ID NO: 2 and 11 of WO2015038958 or SEQ ID NO: 132 and 131
respectively herein), PHP.B (SEQ ID NO: 8 and 9 of WO2015038958 or
SEQ ID NO: 1 and 2 herein), G2B-13 (SEQ ID NO: 12 of WO2015038958
or SEQ ID NO: 3 herein), G2B-26 (SEQ ID NO: 13 of WO2015038958 or
SEQ ID NO: 1 herein), TH1.1-32 (SEQ ID NO: 14 of WO2015038958 or
SEQ ID NO: 4 herein), TH1.1-35 (SEQ ID NO: 15 of WO2015038958 or
SEQ ID NO: 5 herein) or variants thereof. Further, any of the
targeting peptides or amino acid inserts described in WO2015038958,
may be inserted into any parent AAV serotype, such as, but not
limited to, AAV9 (SEQ ID NO: 131 for the DNA sequence and SEQ ID
NO: 132 for the amino acid sequence). In one embodiment, the amino
acid insert is inserted between amino acids 586-592 of the parent
AAV (e.g., AAV9). In another embodiment, the amino acid insert is
inserted between amino acids 588-589 of the parent AAV sequence.
The amino acid insert may be, but is not limited to, any of the
following amino acid sequences, TLAVPFK (SEQ ID NO: 1 of
WO2015038958; herein SEQ ID NO: 876), KFPVALT (SEQ ID NO: 3 of
WO2015038958; herein SEQ ID NO: 877), LAVPFK (SEQ ID NO: 31 of
WO2015038958; herein SEQ ID NO: 878), AVPFK (SEQ ID NO: 32 of
WO2015038958; herein SEQ ID NO: 879), VPFK (SEQ ID NO: 33 of
WO2015038958; herein SEQ ID NO: 880), TLAVPF (SEQ ID NO: 34 of
WO2015038958; herein SEQ ID NO: 881), TLAVP (SEQ ID NO: 35 of
WO2015038958; herein SEQ ID NO: 882), TLAV (SEQ ID NO: 36 of
WO2015038958; herein SEQ ID NO: 883), SVSKPFL (SEQ ID NO: 28 of
WO2015038958; herein SEQ ID NO: 884), FTLTTPK (SEQ ID NO: 29 of
WO2015038958; herein SEQ ID NO: 885), MNATKNV (SEQ ID NO: 30 of
WO2015038958; herein SEQ ID NO: 886), QSSQTPR (SEQ ID NO: 54 of
WO2015038958; herein SEQ ID NO: 887), ILGTGTS (SEQ ID NO: 55 of
WO2015038958; herein SEQ ID NO: 888), TRTNPEA (SEQ ID NO: 56 of
WO2015038958; herein SEQ ID NO: 889), NGGTSSS (SEQ ID NO: 58 of
WO2015038958; herein SEQ ID NO: 890), or YTLSQGW (SEQ ID NO: 60 of
WO2015038958; herein SEQ ID NO: 891). Non-limiting examples of
nucleotide sequences that may encode the amino acid inserts include
the following, AAGTTTCCTGTGGCGTTGACT (for SEQ ID NO: 3 of
WO2015038958; herein SEQ ID NO: 892), ACTTTGGCGGTGCCTTTTAAG (SEQ ID
NO: 24 and 49 of WO2015038958; herein SEQ ID NO: 893),
AGTGTGAGTAAGCCTTTTTTG (SEQ ID NO: 25 of WO2015038958; herein SEQ ID
NO: 894), TTTACGTTGACGACGCCTAAG (SEQ ID NO: 26 of WO2015038958;
herein SEQ ID NO: 895), ATGAATGCTACGAAGAATGTG (SEQ ID NO: 27 of
WO2015038958; herein SEQ ID NO: 896), CAGTCGTCGCAGACGCCTAGG (SEQ ID
NO: 48 of WO2015038958; herein SEQ ID NO: 897),
ATTCTGGGGACTGGTACTTCG (SEQ ID NO: 50 and 52 of WO2015038958; herein
SEQ ID NO: 898), ACGCGGACTAATCCTGAGGCT (SEQ ID NO: 51 of
WO2015038958; herein SEQ ID NO: 899), AATGGGGGGACTAGTAGTTCT (SEQ ID
NO: 53 of WO2015038958; herein SEQ ID NO: 900), or
TATACTTTGTCGCAGGGTTGG (SEQ ID NO: 59 of WO2015038958; herein SEQ ID
NO: 901).
[0069] In one embodiment, the AAV serotype may be, or may have a
sequence as described in International Patent Publication
WO2017100671, the contents of which are herein incorporated by
reference in their entirety, such as, but not limited to, AAV9 (SEQ
ID NO: 45 of WO2017100671, herein SEQ ID NO: 875), PHP.N (SEQ ID
NO: 46 of WO2017100671, herein SEQ ID NO: 873), PHP.S (SEQ ID NO:
47 of WO2017100671, herein SEQ ID NO: 874), or variants thereof.
Further, any of the targeting peptides or amino acid inserts
described in WO2017100671 may be inserted into any parent AAV
serotype, such as, but not limited to, AAV9 (SEQ ID NO: 127 or SEQ
ID NO: 875). In one embodiment, the amino acid insert is inserted
between amino acids 586-592 of the parent AAV (e.g., AAV9). In
another embodiment, the amino acid insert is inserted between amino
acids 588-589 of the parent AAV sequence. The amino acid insert may
be, but is not limited to, any of the following amino acid
sequences, AQTLAVPFKAQ (SEQ ID NO: 1 of WO2017100671; herein SEQ ID
NO: 902), AQSVSKPFLAQ (SEQ ID NO: 2 of WO2017100671; herein SEQ ID
NO: 903), AQFTLTTPKAQ (SEQ ID NO: 3 in the sequence listing of
WO2017100671; herein SEQ ID NO: 904), DGTLAVPFKAQ (SEQ ID NO: 4 in
the sequence listing of WO2017100671; herein SEQ ID NO: 905),
ESTLAVPFKAQ (SEQ ID NO: 5 of WO2017100671; herein SEQ ID NO: 906),
GGTLAVPFKAQ (SEQ ID NO: 6 of WO2017100671; herein SEQ ID NO: 907),
AQTLATPFKAQ (SEQ ID NO: 7 and 33 of WO2017100671; herein SEQ ID NO:
908), ATTLATPFKAQ (SEQ ID NO: 8 of WO2017100671; herein SEQ ID NO:
909), DGTLATPFKAQ (SEQ ID NO: 9 of WO2017100671; herein SEQ ID NO:
910), GGTLATPFKAQ (SEQ ID NO: 10 of WO2017100671; herein SEQ ID NO:
911), SGSLAVPFKAQ (SEQ ID NO: 11 of WO2017100671; herein SEQ ID NO:
912), AQTLAQPFKAQ (SEQ ID NO: 12 of WO2017100671; herein SEQ ID NO:
913), AQTLQQPFKAQ (SEQ ID NO: 13 of WO2017100671; herein SEQ ID NO:
914), AQTLSNPFKAQ (SEQ ID NO: 14 of WO2017100671; herein SEQ ID NO:
915), AQTLAVPFSNP (SEQ ID NO: 15 of WO2017100671; herein SEQ ID NO:
916), QGTLAVPFKAQ (SEQ ID NO: 16 of WO2017100671; herein SEQ ID NO:
917), NQTLAVPFKAQ (SEQ ID NO: 17 of WO2017100671; herein SEQ ID NO:
918), EGSLAVPFKAQ (SEQ ID NO: 18 of WO2017100671; herein SEQ ID NO:
919), SGNLAVPFKAQ (SEQ ID NO: 19 of WO2017100671; herein SEQ ID NO:
920), EGTLAVPFKAQ (SEQ ID NO: 20 of WO2017100671; herein SEQ ID NO:
921), DSTLAVPFKAQ (SEQ ID NO: 21 in Table 1 of WO2017100671; herein
SEQ ID NO: 922), AVTLAVPFKAQ (SEQ ID NO: 22 of WO2017100671; herein
SEQ ID NO: 923), AQTLSTPFKAQ (SEQ ID NO: 23 of WO2017100671; herein
SEQ ID NO: 924), AQTLPQPFKAQ (SEQ ID NO: 24 and 32 of WO2017100671;
herein SEQ ID NO: 925), AQTLSQPFKAQ (SEQ ID NO: 25 of WO2017100671;
herein SEQ ID NO: 926), AQTLQLPFKAQ (SEQ ID NO: 26 of WO2017100671;
herein SEQ ID NO: 927), AQTLTMPFKAQ (SEQ ID NO: 27, and 34 of
WO2017100671 and SEQ ID NO: 35 in the sequence listing of
WO2017100671; herein SEQ ID NO: 928), AQTLTTPFKAQ (SEQ ID NO: 28 of
WO2017100671; herein SEQ ID NO: 929), AQYTLSQGWAQ (SEQ ID NO: 29 of
WO2017100671; herein SEQ ID NO: 930), AQMNATKNVAQ (SEQ ID NO: 30 of
WO2017100671; herein SEQ ID NO: 931), AQVSGGHHSAQ (SEQ ID NO: 31 of
WO2017100671; herein SEQ ID NO: 932), AQTLTAPFKAQ (SEQ ID NO: 35 in
Table 1 of WO2017100671; herein SEQ ID NO: 933), AQTLSKPFKAQ (SEQ
ID NO: 36 of WO2017100671; herein SEQ ID NO: 934), QAVRTSL (SEQ ID
NO: 37 of WO2017100671; herein SEQ ID NO: 935), YTLSQGW (SEQ ID NO:
38 of WO2017100671; herein SEQ ID NO: 891), LAKERLS (SEQ ID NO: 39
of WO2017100671; herein SEQ ID NO: 936), TLAVPFK (SEQ ID NO: 40 in
the sequence listing of WO2017100671; herein SEQ ID NO: 876),
SVSKPFL (SEQ ID NO: 41 of WO2017100671; herein SEQ ID NO: 884),
FTLTTPK (SEQ ID NO: 42 of WO2017100671; herein SEQ ID NO: 885),
MNSTKNV (SEQ ID NO: 43 of WO2017100671; herein SEQ ID NO: 937),
VSGGHHS (SEQ ID NO: 44 of WO2017100671; herein SEQ ID NO: 938),
SAQTLAVPFKAQAQ (SEQ ID NO: 48 of WO2017100671; herein SEQ ID NO:
939), SXXXLAVPFKAQAQ (SEQ ID NO: 49 of WO2017100671 wherein X may
be any amino acid; herein SEQ ID NO: 940), SAQXXXVPFKAQAQ (SEQ ID
NO: 50 of WO2017100671 wherein X may be any amino acid; herein SEQ
ID NO: 941), SAQTLXXXFKAQAQ (SEQ ID NO: 51 of WO2017100671 wherein
X may be any amino acid; herein SEQ ID NO: 942), SAQTLAVXXXAQAQ
(SEQ ID NO: 52 of WO2017100671 wherein X may be any amino acid;
herein SEQ ID NO: 943), SAQTLAVPFXXXAQ (SEQ ID NO: 53 of
WO2017100671 wherein X may be any amino acid; herein SEQ ID NO:
944), TNHQSAQ (SEQ ID NO: 65 of WO2017100671; herein SEQ ID NO:
945), AQAQTGW (SEQ ID NO: 66 of WO2017100671; herein SEQ ID NO:
946), DGTLATPFK (SEQ ID NO: 67 of WO2017100671; herein SEQ ID NO:
947), DGTLATPFKXX (SEQ ID NO: 68 of WO2017100671 wherein X may be
any amino acid; herein SEQ ID NO: 948), LAVPFKAQ (SEQ ID NO: 80 of
WO2017100671; herein SEQ ID NO: 949), VPFKAQ (SEQ ID NO: 81 of
WO2017100671; herein SEQ ID NO: 950), FKAQ (SEQ ID NO: 82 of
WO2017100671; herein SEQ ID NO: 951), AQTLAV (SEQ ID NO: 83 of
WO2017100671; herein SEQ ID NO: 952), AQTLAVPF (SEQ ID NO: 84 of
WO2017100671; herein SEQ ID NO: 953), QAVR (SEQ ID NO: 85 of
WO2017100671; herein SEQ ID NO: 954), AVRT (SEQ ID NO: 86 of
WO2017100671; herein SEQ ID NO: 955), VRTS (SEQ ID NO: 87 of
WO2017100671; herein SEQ ID NO: 956), RTSL (SEQ ID NO: 88 of
WO2017100671; herein SEQ ID NO: 957), QAVRT (SEQ ID NO: 89 of
WO2017100671; herein SEQ ID NO: 958), AVRTS (SEQ ID NO: 90 of
WO2017100671; herein SEQ ID NO: 959), VRTSL (SEQ ID NO: 91 of
WO2017100671; herein SEQ ID NO: 960), QAVRTS (SEQ ID NO: 92 of
WO2017100671; herein SEQ ID NO: 961), or AVRTSL (SEQ ID NO: 93 of
WO2017100671; herein SEQ ID NO: 962).
[0070] Non-limiting examples of nucleotide sequences that may
encode the amino acid inserts include the following,
GATGGGACTTTGGCGGTGCCTTTTAAGGCACAG (SEQ ID NO: 54 of WO2017100671;
herein SEQ ID NO: 963), GATGGGACGTTGGCGGTGCCTTTTAAGGCACAG (SEQ ID
NO: 55 of WO2017100671; herein SEQ ID NO: 964),
CAGGCGGTTAGGACGTCTTTG (SEQ ID NO: 56 of WO2017100671; herein SEQ ID
NO: 965), CAGGTCTTCACGGACTCAGACTATCAG (SEQ ID NO: 57 and 78 of
WO2017100671; herein SEQ ID NO: 966),
CAAGTAAAACCTCTACAAATGTGGTAAAATCG (SEQ ID NO: 58 of WO2017100671;
herein SEQ ID NO: 967), ACTCATCGACCAATACTTGTACTATCTCTCTAGAAC (SEQ
ID NO: 59 of WO2017100671; herein SEQ ID NO: 968),
GGAAGTATTCCTTGGTTTTGAACCCA (SEQ ID NO: 60 of WO2017100671; herein
SEQ ID NO: 969), GGTCGCGGTTCTTGTTTGTGGAT (SEQ ID NO: 61 of
WO2017100671; herein SEQ ID NO: 970), CGACCTTGAAGCGCATGAACTCCT (SEQ
ID NO: 62 of WO2017100671; herein SEQ ID NO: 971),
GTATTCCTTGGTTTTGAACCCAACCGGTCTGCGCCTGTGCMNNMNNMNNMNNMNNMN
NMNNTTGGGCACTCTGGTGGTTTGTC (SEQ ID NO: 63 of WO2017100671 wherein N
may be A, C, T, or G; herein SEQ ID NO: 972), GTATTCC
TTGGTTTTGAACCCAACCGGTC TGCGCMNNMNNMNNAAAAGGCAC CGCCAAA GTTTG (SEQ
ID NO: 69 of WO2017100671 wherein N may be A, C, T, or G; herein
SEQ ID NO: 973),
GTATTCCTTGGTTTTGAACCCAACCGGTCTGCGCCTGTGCMNNMNNMNNCACCGCCAAAG
TTTGGGCACT (SEQ ID NO: 70 of WO2017100671 wherein N may be A, C, T,
or G; herein SEQ ID NO: 974),
GTATTCCTTGGTTTTGAACCCAACCGGTCTGCGCCTGTGCCTTAAAMNNMNNMNNCAA
AGTTTGGGCACTCTGGTGG (SEQ ID NO: 71 of WO2017100671 wherein N may be
A, C, T, or G; herein SEQ ID NO: 975),
GTATTCCTTGGTTTTGAACCCAACCGGTCTGCGCCTGTGCCTTAAAAGGCACMNNMNN
MNNTTGGGCACTCTGGTGGTTTGTG (SEQ ID NO: 72 of WO2017100671 wherein N
may be A, C, T, or G; herein SEQ ID NO: 976), ACTTTGGCGGTGCCTTTTAAG
(SEQ ID NO: 74 of WO2017100671; herein SEQ ID NO: 893),
AGTGTGAGTAAGCCTTTTTTG (SEQ ID NO: 75 of WO2017100671; herein SEQ ID
NO: 894), TTTACGTTGACGACGCCTAAG (SEQ ID NO: 76 of WO2017100671;
herein SEQ ID NO: 895), TATACTTTGTCGCAGGGTTGG (SEQ ID NO: 77 of
WO2017100671; herein SEQ ID NO: 901), or CTTGCGAAGGAGCGGCTTTCG (SEQ
ID NO: 79 of WO2017100671; herein SEQ ID NO: 977).
[0071] In one embodiment, the AAV serotype may be, or may have a
sequence as described in U.S. Pat. No. 9,624,274, the contents of
which are herein incorporated by reference in their entirety, such
as, but not limited to, AAV1 (SEQ ID NO: 181 of U.S. Pat. No.
9,624,274), AAV6 (SEQ ID NO: 182 of U.S. Pat. No. 9,624,274), AAV2
(SEQ ID NO: 183 of U.S. Pat. No. 9,624,274), AAV3b (SEQ ID NO: 184
of U.S. Pat. No. 9,624,274), AAV7 (SEQ ID NO: 185 of U.S. Pat. No.
9,624,274), AAV8 (SEQ ID NO: 186 of U.S. Pat. No. 9,624,274), AAV10
(SEQ ID NO: 187 of U.S. Pat. No. 9,624,274), AAV4 (SEQ ID NO: 188
of U.S. Pat. No. 9,624,274), AAV11 (SEQ ID NO: 189 of U.S. Pat. No.
9,624,274), bAAV (SEQ ID NO: 190 of U.S. Pat. No. 9,624,274), AAV5
(SEQ ID NO: 191 of U.S. Pat. No. 9,624,274), GPV (SEQ ID NO: 192 of
U.S. Pat. No. 9,624,274; herein SEQ ID NO: 992), B19 (SEQ ID NO:
193 of U.S. Pat. No. 9,624,274; herein SEQ ID NO: 993), MVM (SEQ ID
NO: 194 of U.S. Pat. No. 9,624,274; herein SEQ ID NO: 994), FPV
(SEQ ID NO: 195 of U.S. Pat. No. 9,624,274; herein SEQ ID NO: 995),
CPV (SEQ ID NO: 196 of U.S. Pat. No. 9,624,274; herein SEQ ID NO:
996) or variants thereof. Further, any of the structural protein
inserts described in U.S. Pat. No. 9,624,274, may be inserted into,
but not limited to, 1-453 and 1-587 of any parent AAV serotype,
such as, but not limited to, AAV2 (SEQ ID NO: 183 of U.S. Pat. No.
9,624,274). The amino acid insert may be, but is not limited to,
any of the following amino acid sequences, VNLTWSRASG (SEQ ID NO:
50 of U.S. Pat. No. 9,624,274; herein SEQ ID NO: 1375),
EFCINHRGYWVCGD (SEQ ID NO:55 of U.S. Pat. No. 9,624,274; herein SEQ
ID NO: 1376), EDGQVMDVDLS (SEQ ID NO: 85 of U.S. Pat. No.
9,624,274; herein SEQ ID NO: 1377), EKQRNGTLT (SEQ ID NO: 86 of
U.S. Pat. No. 9,624,274; herein SEQ ID NO: 1378), TYQCRVTHPHLPRALMR
(SEQ ID NO: 87 of U.S. Pat. No. 9,624,274; herein SEQ ID NO: 1379),
RHSTTQPRKTKGSG (SEQ ID NO: 88 of U.S. Pat. No. 9,624,274; herein
SEQ ID NO: 1380), DSNPRGVSAYLSR (SEQ ID NO: 89 of U.S. Pat. No.
9,624,274; herein SEQ ID NO: 1381), TITCLWDLAPSK (SEQ ID NO: 90 of
U.S. Pat. No. 9,624,274; herein SEQ ID NO: 1382), KTKGSGFFVF (SEQ
ID NO: 91 of U.S. Pat. No. 9,624,274; herein SEQ ID NO: 1383),
THPHLPRALMRS (SEQ ID NO: 92 of U.S. Pat. No. 9,624,274; herein SEQ
ID NO: 1384), GETYQCRVTHPHLPRALMRSTTK (SEQ ID NO: 93 of U.S. Pat.
No. 9,624,274; herein SEQ ID NO: 1385), LPRALMRS (SEQ ID NO: 94 of
U.S. Pat. No. 9,624,274; herein SEQ ID NO: 1386), INHRGYWV (SEQ ID
NO: 95 of U.S. Pat. No. 9,624,274; herein SEQ ID NO: 1387),
CDAGSVRTNAPD (SEQ ID NO: 60 of U.S. Pat. No. 9,624,274; herein SEQ
ID NO: 1388), AKAVSNLTESRSESLQS (SEQ ID NO: 96 of U.S. Pat. No.
9,624,274; herein SEQ ID NO: 1389), SLTGDEFKKVLET (SEQ ID NO: 97 of
U.S. Pat. No. 9,624,274; herein SEQ ID NO: 1390), REAVAYRFEED (SEQ
ID NO: 98 of U.S. Pat. No. 9,624,274; herein SEQ ID NO: 1391),
INPEIITLDG (SEQ ID NO: 99 of U.S. Pat. No. 9,624,274; herein SEQ ID
NO: 1392), DISVTGAPVITATYL (SEQ ID NO: 100 of U.S. Pat. No.
9,624,274; herein SEQ ID NO: 1393), DISVTGAPVITA (SEQ ID NO: 101 of
U.S. Pat. No. 9,624,274; herein SEQ ID NO: 1394), PKTVSNLTESSSESVQS
(SEQ ID NO: 102 of U.S. Pat. No. 9,624,274; herein SEQ ID NO:
1395), SLMGDEFKAVLET (SEQ ID NO: 103 of U.S. Pat. No. 9,624,274;
herein SEQ ID NO: 1396), QHSVAYTFEED (SEQ ID NO: 104 of U.S. Pat.
No. 9,624,274; herein SEQ ID NO: 1397), INPEIITRDG (SEQ ID NO: 105
of U.S. Pat. No. 9,624,274; herein SEQ ID NO: 1398),
DISLTGDPVITASYL (SEQ ID NO: 106 of U.S. Pat. No. 9,624,274; herein
SEQ ID NO: 1399), DISLTGDPVITA (SEQ ID NO: 107 of U.S. Pat. No.
9,624,274; herein SEQ ID NO: 1400), DQSIDFEIDSA (SEQ ID NO: 108 of
U.S. Pat. No. 9,624,274; herein SEQ ID NO: 1401),
KNVSEDLPLPTFSPTLLGDS (SEQ ID NO: 109 of U.S. Pat. No. 9,624,274;
herein SEQ ID NO: 1402), KNVSEDLPLPT (SEQ ID NO: 110 of U.S. Pat.
No. 9,624,274; herein SEQ ID NO: 1403), CDSGRVRTDAPD (SEQ ID NO:
111 of U.S. Pat. No. 9,624,274; herein SEQ ID NO: 1404),
FPEHLLVDFLQSLS (SEQ ID NO: 112 of U.S. Pat. No. 9,624,274; herein
SEQ ID NO: 1405), DAEFRHDSG (SEQ ID NO: 65 of U.S. Pat. No.
9,624,274; herein SEQ ID NO: 1406), HYAAAQWDFGNTMCQL (SEQ ID NO:
113 of U.S. Pat. No. 9,624,274; herein SEQ ID NO: 1407),
YAAQWDFGNTMCQ (SEQ ID NO: 114 of U.S. Pat. No. 9,624,274; herein
SEQ ID NO: 1408), RSQKEGLHYT (SEQ ID NO: 115 of U.S. Pat. No.
9,624,274; herein SEQ ID NO: 1409), SSRTPSDKPVAHWANPQAE (SEQ ID NO:
116 of U.S. Pat. No. 9,624,274; herein SEQ ID NO: 1410),
SRTPSDKPVAHWANP (SEQ ID NO: 117 of U.S. Pat. No. 9,624,274; herein
SEQ ID NO: 1411), SSRTPSDKP (SEQ ID NO: 118 of U.S. Pat. No.
9,624,274; herein SEQ ID NO: 1412), NADGNVDYHMNSVP (SEQ ID NO: 119
of U.S. Pat. No. 9,624,274; herein SEQ ID NO: 1413), DGNVDYHMNSV
(SEQ ID NO: 120 of U.S. Pat. No. 9,624,274; herein SEQ ID NO:
1414), RSFKEFLQSSLRALRQ (SEQ ID NO: 121 of U.S. Pat. No. 9,624,274;
herein SEQ ID NO: 1415); FKEFLQSSLRA (SEQ ID NO: 122 of U.S. Pat.
No. 9,624,274; herein SEQ ID NO: 1416), or QMWAPQWGPD (SEQ ID NO:
123 of U.S. Pat. No. 9,624,274; herein SEQ ID NO: 1417).
[0072] In one embodiment, the AAV serotype may be, or may have a
sequence as described in U.S. Pat. No. 9,475,845, the contents of
which are herein incorporated by reference in their entirety, such
as, but not limited to, AAV capsid proteins comprising modification
of one or more amino acids at amino acid positions 585 to 590 of
the native AAV2 capsid protein. Further the modification may result
in, but not limited to, the amino acid sequence RGNRQA (SEQ ID NO:
3 of U.S. Pat. No. 9,475,845; herein SEQ ID NO: 1418), SSSTDP (SEQ
ID NO: 4 of U.S. Pat. No. 9,475,845; herein SEQ ID NO: 1419),
SSNTAP (SEQ ID NO: 5 of U.S. Pat. No. 9,475,845; herein SEQ ID NO:
1420), SNSNLP (SEQ ID NO: 6 of U.S. Pat. No. 9,475,845; herein SEQ
ID NO: 1421), SSTTAP (SEQ ID NO: 7 of U.S. Pat. No. 9,475,845;
herein SEQ ID NO: 1422), AANTAA (SEQ ID NO: 8 of U.S. Pat. No.
9,475,845; herein SEQ ID NO: 1423), QQNTAP (SEQ ID NO: 9 of U.S.
Pat. No. 9,475,845; herein SEQ ID NO: 1424), SAQAQA (SEQ ID NO: 10
of U.S. Pat. No. 9,475,845; herein SEQ ID NO: 1425), QANTGP (SEQ ID
NO: 11 of U.S. Pat. No. 9,475,845; herein SEQ ID NO: 1426), NATTAP
(SEQ ID NO: 12 of U.S. Pat. No. 9,475,845; herein SEQ ID NO: 1427),
SSTAGP (SEQ ID NO: 13 and 20 of U.S. Pat. No. 9,475,845; herein SEQ
ID NO: 1428), QQNTAA (SEQ ID NO: 14 of U.S. Pat. No. 9,475,845;
herein SEQ ID NO: 1429), PSTAGP (SEQ ID NO: 15 of U.S. Pat. No.
9,475,845; herein SEQ ID NO: 1430), NQNTAP (SEQ ID NO: 16 of U.S.
Pat. No. 9,475,845; herein SEQ ID NO: 1431), QAANAP (SEQ ID NO: 17
of U.S. Pat. No. 9,475,845; herein SEQ ID NO: 1432), SIVGLP (SEQ ID
NO: 18 of U.S. Pat. No. 9,475,845; herein SEQ ID NO: 1433), AASTAA
(SEQ ID NO: 19, and 27 of U.S. Pat. No. 9,475,845; herein SEQ ID
NO: 1434), SQNTTA (SEQ ID NO: 21 of U.S. Pat. No. 9,475,845; herein
SEQ ID NO: 1435), QQDTAP (SEQ ID NO: 22 of U.S. Pat. No. 9,475,845;
herein SEQ ID NO: 1436), QTNTGP (SEQ ID NO: 23 of U.S. Pat. No.
9,475,845; herein SEQ ID NO: 1437), QTNGAP (SEQ ID NO: 24 of U.S.
Pat. No. 9,475,845; herein SEQ ID NO: 1438), QQNAAP (SEQ ID NO: 25
of U.S. Pat. No. 9,475,845; herein SEQ ID NO: 1439), or AANTQA (SEQ
ID NO: 26 of U.S. Pat. No. 9,475,845; herein SEQ ID NO: 1440). In
one embodiment, the amino acid modification is a substitution at
amino acid positions 262 through 265 in the native AAV2 capsid
protein or the corresponding position in the capsid protein of
another AAV with a targeting sequence. The targeting sequence may
be, but is not limited to, any of the amino acid sequences, NGRAHA
(SEQ ID NO: 38 of U.S. Pat. No. 9,475,845; herein SEQ ID NO: 1441),
QPEHSST (SEQ ID NO: 39 and 50 of U.S. Pat. No. 9,475,845; herein
SEQ ID NO: 1442), VNTANST (SEQ ID NO: 40 of U.S. Pat. No.
9,475,845; herein SEQ ID NO: 1443), HGPMQKS (SEQ ID NO: 41 of
US9475845; herein SEQ ID NO: 1444), PHKPPLA (SEQ ID NO: 42 of U.S.
Pat. No. 9,475,845; herein SEQ ID NO: 1445), IKNNEMW (SEQ ID NO: 43
of U.S. Pat. No. 9,475,845; herein SEQ ID NO: 1446), RNLDTPM (SEQ
ID NO: 44 of U.S. Pat. No. 9,475,845; herein SEQ ID NO: 1447),
VDSHRQS (SEQ ID NO: 45 of U.S. Pat. No. 9,475,845; herein SEQ ID
NO: 1448), YDSKTKT (SEQ ID NO: 46 of U.S. Pat. No. 9,475,845;
herein SEQ ID NO: 1449), SQLPHQK (SEQ ID NO: 47 of U.S. Pat. No.
9,475,845; herein SEQ ID NO: 1450), STMQQNT (SEQ ID NO: 48 of U.S.
Pat. No. 9,475,845; herein SEQ ID NO: 1451), TERYMTQ (SEQ ID NO: 49
of U.S. Pat. No. 9,475,845; herein SEQ ID NO: 1452), DASLSTS (SEQ
ID NO: 51 of U.S. Pat. No. 9,475,845; herein SEQ ID NO: 1453),
DLPNKKT (SEQ ID NO: 52 of U.S. Pat. No. 9,475,845; herein SEQ ID
NO: 1454), DLTAARL (SEQ ID NO: 53 of U.S. Pat. No. 9,475,845;
herein SEQ ID NO: 1455), EPHQFNY (SEQ ID NO: 54 of U.S. Pat. No.
9,475,845; herein SEQ ID NO: 1456), EPQSNHT (SEQ ID NO: 55 of U.S.
Pat. No. 9,475,845; herein SEQ ID NO: 1457), MSSWPSQ (SEQ ID NO: 56
of U.S. Pat. No. 9,475,845; herein SEQ ID NO: 1458), NPKHNAT (SEQ
ID NO: 57 of U.S. Pat. No. 9,475,845; herein SEQ ID NO: 1459),
PDGMRTT (SEQ ID NO: 58 of U.S. Pat. No. 9,475,845; herein SEQ ID
NO: 1460), PNNNKTT (SEQ ID NO: 59 of U.S. Pat. No. 9,475,845;
herein SEQ ID NO: 1461), QSTTHDS (SEQ ID NO: 60 of U.S. Pat. No.
9,475,845; herein SEQ ID NO: 1462), TGSKQKQ (SEQ ID NO: 61 of U.S.
Pat. No. 9,475,845; herein SEQ ID NO: 1463), SLKHQAL (SEQ ID NO: 62
of U.S. Pat. No. 9,475,845; herein SEQ ID NO: 1464), SPIDGEQ (SEQ
ID NO: 63 of U.S. Pat. No. 9,475,845; herein SEQ ID NO: 1465),
WIFPWIQL (SEQ ID NO: 64 and 112 of U.S. Pat. No. 9,475,845; herein
SEQ ID NO: 1466), CDCRGDCFC (SEQ ID NO: 65 of U.S. Pat. No.
9,475,845; herein SEQ ID NO: 1467), CNGRC (SEQ ID NO: 66 of U.S.
Pat. No. 9,475,845; herein SEQ ID NO: 1468), CPRECES (SEQ ID NO: 67
of U.S. Pat. No. 9,475,845; herein SEQ ID NO: 1469), CTTHWGFTLC
(SEQ ID NO: 68 and 123 of U.S. Pat. No. 9,475,845; herein SEQ ID
NO: 1470), CGRRAGGSC (SEQ ID NO: 69 of U.S. Pat. No. 9,475,845;
herein SEQ ID NO: 1471), CKGGRAKDC (SEQ ID NO: 70 of U.S. Pat. No.
9,475,845; herein SEQ ID NO: 1472), CVPELGHEC (SEQ ID NO: 71 and
115 of U.S. Pat. No. 9,475,845; herein SEQ ID NO: 1473), CRRETAWAK
(SEQ ID NO: 72 of U.S. Pat. No. 9,475,845; herein SEQ ID NO: 1474),
VSWFSHRYSPFAVS (SEQ ID NO: 73 of U.S. Pat. No. 9,475,845; herein
SEQ ID NO: 1475), GYRDGYAGPILYN (SEQ ID NO: 74 of U.S. Pat. No.
9,475,845; herein SEQ ID NO: 1476), XXXYXXX (SEQ ID NO: 75 of U.S.
Pat. No. 9,475,845; herein SEQ ID NO: 1477), YXNW (SEQ ID NO: 76 of
U.S. Pat. No. 9,475,845; herein SEQ ID NO: 1478), RPLPPLP (SEQ ID
NO: 77 of U.S. Pat. No. 9,475,845; herein SEQ ID NO: 1479), APPLPPR
(SEQ ID NO: 78 of U.S. Pat. No. 9,475,845; herein SEQ ID NO: 1480),
DVFYPYPYASGS (SEQ ID NO: 79 of U.S. Pat. No. 9,475,845; herein SEQ
ID NO: 1481), MYWYPY (SEQ ID NO: 80 of U.S. Pat. No. 9,475,845;
herein SEQ ID NO: 1482), DITWDQLWDLMK (SEQ ID NO: 81 of U.S. Pat.
No. 9,475,845; herein SEQ ID NO: 1483), CWDDXWLC (SEQ ID NO: 82 of
U.S. Pat. No. 9,475,845; herein SEQ ID NO: 1484), EWCEYLGGYLRCYA
(SEQ ID NO: 83 of U.S. Pat. No. 9,475,845; herein SEQ ID NO: 1485),
YXCXXGPXTWXCXP (SEQ ID NO: 84 of U.S. Pat. No. 9,475,845; herein
SEQ ID NO: 1486), IEGPTLRQWLAARA (SEQ ID NO: 85 of U.S. Pat. No.
9,475,845; herein SEQ ID NO: 1487), LWXXX (SEQ ID NO: 86 of
US9475845; herein SEQ ID NO: 1488), XFXXYLW (SEQ ID NO: 87 of U.S.
Pat. No. 9,475,845; herein SEQ ID NO: 1489), SSIISHFRWGLCD (SEQ ID
NO: 88 of U.S. Pat. No. 9,475,845; herein SEQ ID NO: 1490),
MSRPACPPNDKYE (SEQ ID NO: 89 of U.S. Pat. No. 9,475,845; herein SEQ
ID NO: 1491), CLRSGRGC (SEQ ID NO: 90 of U.S. Pat. No. 9,475,845;
herein SEQ ID NO: 1492), CHWMFSPWC (SEQ ID NO: 91 of U.S. Pat. No.
9,475,845; herein SEQ ID NO: 1493), WXXF (SEQ ID NO: 92 of U.S.
Pat. No. 9,475,845; herein SEQ ID NO: 1494), CSSRLDAC (SEQ ID NO:
93 of U.S. Pat. No. 9,475,845; herein SEQ ID NO: 1495), CLPVASC
(SEQ ID NO: 94 of U.S. Pat. No. 9,475,845; herein SEQ ID NO: 1496),
CGFECVRQCPERC (SEQ ID NO: 95 of U.S. Pat. No. 9,475,845; herein SEQ
ID NO: 1497), CVALCREACGEGC (SEQ ID NO: 96 of U.S. Pat. No.
9,475,845; herein SEQ ID NO: 1498), SWCEPGWCR (SEQ ID NO: 97 of
U.S. Pat. No. 9,475,845; herein SEQ ID NO: 1499), YSGKWGW (SEQ ID
NO: 98 of U.S. Pat. No. 9,475,845; herein SEQ ID NO: 1500), GLSGGRS
(SEQ ID NO: 99 of U.S. Pat. No. 9,475,845; herein SEQ ID NO: 1501),
LMLPRAD (SEQ ID NO: 100 of U.S. Pat. No. 9,475,845; herein SEQ ID
NO: 1502), CSCFRDVCC (SEQ ID NO: 101 of U.S. Pat. No. 9,475,845;
herein SEQ ID NO: 1503), CRDVVSVIC (SEQ ID NO: 102 of U.S. Pat. No.
9,475,845; herein SEQ ID NO: 1504), MARSGL (SEQ ID NO: 103 of U.S.
Pat. No. 9,475,845; herein SEQ ID NO: 1505), MARAKE (SEQ ID NO: 104
of U.S. Pat. No. 9,475,845; herein SEQ ID NO: 1506), MSRTMS (SEQ ID
NO: 105 of U.S. Pat. No. 9,475,845; herein SEQ ID NO: 1507), KCCYSL
(SEQ ID NO: 106 of U.S. Pat. No. 9,475,845; herein SEQ ID NO:
1508), MYWGDSHWLQYWYE (SEQ ID NO: 107 of U.S. Pat. No. 9,475,845;
herein SEQ ID NO: 1509), MQLPLAT (SEQ ID NO: 108 of U.S. Pat. No.
9,475,845; herein SEQ ID NO: 1510), EWLS (SEQ ID NO: 109 of U.S.
Pat. No. 9,475,845; herein SEQ ID NO: 1511), SNEW (SEQ ID NO: 110
of U.S. Pat. No. 9,475,845; herein SEQ ID NO: 1512), TNYL (SEQ ID
NO: 111 of U.S. Pat. No. 9,475,845; herein SEQ ID NO: 1513),
WDLAWMFRLPVG (SEQ ID NO: 113 of U.S. Pat. No. 9,475,845; herein SEQ
ID NO: 1514), CTVALPGGYVRVC (SEQ ID NO: 114 of U.S. Pat. No.
9,475,845; herein SEQ ID NO: 1515), CVAYCIEHHCWTC (SEQ ID NO: 116
of U.S. Pat. No. 9,475,845; herein SEQ ID NO: 1516), CVFAHNYDYLVC
(SEQ ID NO: 117 of U.S. Pat. No. 9,475,845; herein SEQ ID NO:
1517), CVFTSNYAFC (SEQ ID NO: 118 of U.S. Pat. No. 9,475,845;
herein SEQ ID NO: 1518), VHSPNKK (SEQ ID NO: 119 of U.S. Pat. No.
9,475,845; herein SEQ ID NO: 1519), CRGDGWC (SEQ ID NO: 120 of U.S.
Pat. No. 9,475,845; herein SEQ ID NO: 1520), XRGCDX (SEQ ID NO: 121
of U.S. Pat. No. 9,475,845; herein SEQ ID NO: 1521), PXXX (SEQ ID
NO: 122 of U.S. Pat. No. 9,475,845; herein SEQ ID NO: 1522),
SGKGPRQITAL (SEQ ID NO: 124 of U.S. Pat. No. 9,475,845; herein SEQ
ID NO: 1523), AAAAAAAAAXXXXX (SEQ ID NO: 125 of U.S. Pat. No.
9,475,845; herein SEQ ID NO: 1524), VYMSPF (SEQ ID NO: 126 of U.S.
Pat. No. 9,475,845; herein SEQ ID NO: 1525), ATWLPPR (SEQ ID NO:
127 of U.S. Pat. No. 9,475,845; herein SEQ ID NO: 1526),
HTMYYHHYQHHL (SEQ ID NO: 128 of U.S. Pat. No. 9,475,845; herein SEQ
ID NO: 1527), SEVGCRAGPLQWLCEKYFG (SEQ ID NO: 129 of U.S. Pat. No.
9,475,845; herein SEQ ID NO: 1528), CGLLPVGRPDRNVWRWLC (SEQ ID NO:
130 of U.S. Pat. No. 9,475,845; herein SEQ ID NO: 1529),
CKGQCDRFKGLPWEC (SEQ ID NO: 131 of U.S. Pat. No. 9,475,845; herein
SEQ ID NO: 1530), SGRSA (SEQ ID NO: 132 of U.S. Pat. No. 9,475,845;
herein SEQ ID NO: 1531), WGFP (SEQ ID NO: 133 of U.S. Pat. No.
9,475,845; herein SEQ ID NO: 1532), AEPMPHSLNFSQYLWYT (SEQ ID NO:
134 of U.S. Pat. No. 9,475,845; herein SEQ ID NO: 1533), WAYXSP
(SEQ ID NO: 135 of U.S. Pat. No. 9,475,845; herein SEQ ID NO:
1534), IELLQAR (SEQ ID NO: 136 of U.S. Pat. No. 9,475,845; herein
SEQ ID NO: 1535), AYTKCSRQWRTCMTTH (SEQ ID NO: 137 of U.S. Pat. No.
9,475,845; herein SEQ ID NO: 1536), PQNSKIPGPTFLDPH (SEQ ID NO: 138
of U.S. Pat. No. 9,475,845; herein SEQ ID NO: 1537),
SMEPALPDWWWKMFK (SEQ ID NO: 139 of U.S. Pat. No. 9,475,845; herein
SEQ ID NO: 1538), ANTPCGPYTHDCPVKR (SEQ ID NO: 140 of U.S. Pat. No.
9,475,845; herein SEQ ID NO: 1539), TACHQHVRMVRP (SEQ ID NO: 141 of
U.S. Pat. No. 9,475,845; herein SEQ ID NO: 1540), VPWMEPAYQRFL (SEQ
ID NO: 142 of U.S. Pat. No. 9,475,845; herein SEQ ID NO: 1541),
DPRATPGS (SEQ ID NO: 143 of U.S. Pat. No. 9,475,845; herein SEQ ID
NO: 1542), FRPNRAQDYNTN (SEQ ID NO: 144 of U.S. Pat. No. 9,475,845;
herein SEQ ID NO: 1543), CTKNSYLMC (SEQ ID NO: 145 of U.S. Pat. No.
9,475,845; herein SEQ ID NO: 1544), CXXTXXXGXGC (SEQ ID NO: 146 of
U.S. Pat. No. 9,475,845; herein SEQ ID NO: 1545), CPIEDRPMC (SEQ ID
NO: 147 of U.S. Pat. No. 9,475,845; herein SEQ ID NO: 1546),
HEWSYLAPYPWF (SEQ ID NO: 148 of U.S. Pat. No. 9,475,845; herein SEQ
ID NO: 1547), MCPKHPLGC (SEQ ID NO: 149 of U.S. Pat. No. 9,475,845;
herein SEQ ID NO: 1548), RMWPSSTVNLSAGRR (SEQ ID NO: 150 of U.S.
Pat. No. 9,475,845; herein SEQ ID NO: 1549), SAKTAVSQRVWLPSHRGGEP
(SEQ ID NO: 151 of U.S. Pat. No. 9,475,845; herein SEQ ID NO:
1550), KSREHVNNSACPSKRITAAL (SEQ ID NO: 152 of U.S. Pat. No.
9,475,845; herein SEQ ID NO: 1551), EGFR (SEQ ID NO: 153 of
US9475845; herein SEQ ID NO: 1552), AGLGVR (SEQ ID NO: 154 of U.S.
Pat. No. 9,475,845; herein SEQ ID NO: 1553), GTRQGHTMRLGVSDG (SEQ
ID NO: 155 of U.S. Pat. No. 9,475,845; herein SEQ ID NO: 1554),
IAGLATPGWSHWLAL (SEQ ID NO: 156 of U.S. Pat. No. 9,475,845; herein
SEQ ID NO: 1555), SMSIARL (SEQ ID NO: 157 of U.S. Pat. No.
9,475,845; herein SEQ ID NO: 1556), HTFEPGV (SEQ ID NO: 158 of U.S.
Pat. No. 9,475,845; herein SEQ ID NO: 1557), NTSLKRISNKRIRRK (SEQ
ID NO: 159 of US9475845; herein SEQ ID NO: 1558), LRIKRKRRKRKKTRK
(SEQ ID NO: 160 of U.S. Pat. No. 9,475,845; herein SEQ ID NO:
1559), GGG, GFS, LWS, EGG, LLV, LSP, LBS, AGG, GRR, GGH and
GTV.
[0073] In one embodiment, the AAV serotype may be, or may have a
sequence as described in United States Publication No. US
20160369298, the contents of which are herein incorporated by
reference in their entirety, such as, but not limited to,
site-specific mutated capsid protein of AAV2 (SEQ ID NO: 97 of US
20160369298; herein SEQ ID NO: 1560) or variants thereof, wherein
the specific site is at least one site selected from sites R447,
G453, S578, N587, N587+1, S662 of VP1 or fragment thereof.
[0074] Further, any of the mutated sequences described in US
20160369298, may be or may have, but not limited to, any of the
following sequences SDSGASN (SEQ ID NO: 1 and SEQ ID NO: 231 of
US20160369298; herein SEQ ID NO: 1561), SPSGASN (SEQ ID NO: 2 of
US20160369298; herein SEQ ID NO: 1562), SHSGASN (SEQ ID NO: 3 of
US20160369298; herein SEQ ID NO: 1563), SRSGASN (SEQ ID NO: 4 of
US20160369298; herein SEQ ID NO: 1564), SKSGASN (SEQ ID NO: 5 of
US20160369298; herein SEQ ID NO: 1565), SNSGASN (SEQ ID NO: 6 of
US20160369298; herein SEQ ID NO: 1566), SGSGASN (SEQ ID NO: 7 of
US20160369298; herein SEQ ID NO: 1567), SASGASN (SEQ ID NO: 8, 175,
and 221 of US20160369298; herein SEQ ID NO: 1568), SESGTSN (SEQ ID
NO: 9 of US20160369298; herein SEQ ID NO: 1569), STTGGSN (SEQ ID
NO: 10 of US20160369298; herein SEQ ID NO: 1570), SSAGSTN (SEQ ID
NO: 11 of US20160369298; herein SEQ ID NO: 1571), NNDSQA (SEQ ID
NO: 12 of US20160369298; herein SEQ ID NO: 1572), NNRNQA (SEQ ID
NO: 13 of US20160369298; herein SEQ ID NO: 1573), NNNKQA (SEQ ID
NO: 14 of US20160369298; herein SEQ ID NO: 1574), NAKRQA (SEQ ID
NO: 15 of US20160369298; herein SEQ ID NO: 1575), NDEHQA (SEQ ID
NO: 16 of US20160369298; herein SEQ ID NO: 1576), NTSQKA (SEQ ID
NO: 17 of US20160369298; herein SEQ ID NO: 1577),
YYLSRTNTPSGTDTQSRLVFSQAGA (SEQ ID NO: 18 of US20160369298; herein
SEQ ID NO: 1578), YYLSRTNTDSGTETQSGLDFSQAGA (SEQ ID NO: 19 of
US20160369298; herein SEQ ID NO: 1579), YYLSRTNTESGTPTQSALEFSQAGA
(SEQ ID NO: 20 of US20160369298; herein SEQ ID NO: 1580),
YYLSRTNTHSGTHTQSPLHFSQAGA (SEQ ID NO: 21 of US20160369298; herein
SEQ ID NO: 1581), YYLSRTNTSSGTITISHLIFSQAGA (SEQ ID NO: 22 of
US20160369298; herein SEQ ID NO: 1582), YYLSRTNTRSGIMTKSSLMFSQAGA
(SEQ ID NO: 23 of US20160369298; herein SEQ ID NO: 1583),
YYLSRTNTKSGRKTLSNLSFSQAGA (SEQ ID NO: 24 of US20160369298; herein
SEQ ID NO: 1584), YYLSRTNDGSGPVTPSKLRFSQRGA (SEQ ID NO: 25 of
US20160369298; herein SEQ ID NO: 1585), YYLSRTNAASGHATHSDLKFSQPGA
(SEQ ID NO: 26 of US20160369298; herein SEQ ID NO: 1586),
YYLSRTNGQAGSLTMSELGFSQVGA (SEQ ID NO: 27 of US20160369298; herein
SEQ ID NO: 1587), YYLSRTNSTGGNQTTSQLLFSQLSA (SEQ ID NO: 28 of
US20160369298; herein SEQ ID NO: 1588), YFLSRTNNNTGLNTNSTLNFSQGRA
(SEQ ID NO: 29 of US20160369298; herein SEQ ID NO: 1589),
SKTGADNNNSEYSWTG (SEQ ID NO: 30 of US20160369298; herein SEQ ID NO:
1590), SKTDADNNNSEYSWTG (SEQ ID NO: 31 of US20160369298; herein SEQ
ID NO: 1591), SKTEADNNNSEYSWTG (SEQ ID NO: 32 of US20160369298;
herein SEQ ID NO: 1592), SKTPADNNNSEYSWTG (SEQ ID NO: 33 of
US20160369298; herein SEQ ID NO: 1593), SKTHADNNNSEYSWTG (SEQ ID
NO: 34 of US20160369298; herein SEQ ID NO: 1594), SKTQADNNNSEYSWTG
(SEQ ID NO: 35 of US20160369298; herein SEQ ID NO: 1595),
SKTIADNNNSEYSWTG (SEQ ID NO: 36 of US20160369298; herein SEQ ID NO:
1596), SKTMADNNNSEYSWTG (SEQ ID NO: 37 of US20160369298; herein SEQ
ID NO: 1597), SKTRADNNNSEYSWTG (SEQ ID NO: 38 of US20160369298;
herein SEQ ID NO: 1598), SKTNADNNNSEYSWTG (SEQ ID NO: 39 of
US20160369298; herein SEQ ID NO: 1599), SKTVGRNNNSEYSWTG (SEQ ID
NO: 40 of US20160369298; herein SEQ ID NO: 1600), SKTADRNNNSEYSWTG
(SEQ ID NO: 41 of US20160369298; herein SEQ ID NO: 1601),
SKKLSQNNNSKYSWQG (SEQ ID NO: 42 of US20160369298; herein SEQ ID NO:
1602), SKPTTGNNNSDYSWPG (SEQ ID NO: 43 of US20160369298; herein SEQ
ID NO: 1603), STQKNENNNSNYSWPG (SEQ ID NO: 44 of US20160369298;
herein SEQ ID NO: 1604), HKDDEGKF (SEQ ID NO: 45 of US20160369298;
herein SEQ ID NO: 1605), HKDDNRKF (SEQ ID NO: 46 of US20160369298;
herein SEQ ID NO: 1606), HKDDTNKF (SEQ ID NO: 47 of US20160369298;
herein SEQ ID NO: 1607), HEDSDKNF (SEQ ID NO: 48 of US20160369298;
herein SEQ ID NO: 1608), HRDGADSF (SEQ ID NO: 49 of US20160369298;
herein SEQ ID NO: 1609), HGDNKSRF (SEQ ID NO: 50 of US20160369298;
herein SEQ ID NO: 1610), KQGSEKTNVDFEEV (SEQ ID NO: 51 of
US20160369298; herein SEQ ID NO: 1611), KQGSEKTNVDSEEV (SEQ ID NO:
52 of US20160369298; herein SEQ ID NO: 1612), KQGSEKTNVDVEEV (SEQ
ID NO: 53 of US20160369298; herein SEQ ID NO: 1613), KQGSDKTNVDDAGV
(SEQ ID NO: 54 of US20160369298; herein SEQ ID NO: 1614),
KQGSSKTNVDPREV (SEQ ID NO: 55 of US20160369298; herein SEQ ID NO:
1615), KQGSRKTNVDHKQV (SEQ ID NO: 56 of US20160369298; herein SEQ
ID NO: 1616), KQGSKGGNVDTNRV (SEQ ID NO: 57 of US20160369298;
herein SEQ ID NO: 1617), KQGSGEANVDNGDV (SEQ ID NO: 58 of
US20160369298; herein SEQ ID NO: 1618), KQDAAADNIDYDHV (SEQ ID NO:
59 of US20160369298; herein SEQ ID NO: 1619), KQSGTRSNAAASSV (SEQ
ID NO: 60 of US20160369298; herein SEQ ID NO: 1620), KENTNTNDTELTNV
(SEQ ID NO: 61 of US20160369298; herein SEQ ID NO: 1621),
QRGNNVAATADVNT (SEQ ID NO: 62 of US20160369298; herein SEQ ID NO:
1622), QRGNNEAATADVNT (SEQ ID NO: 63 of US20160369298; herein SEQ
ID NO: 1623), QRGNNPAATADVNT (SEQ ID NO: 64 of US20160369298;
herein SEQ ID NO: 1624), QRGNNHAATADVNT (SEQ ID NO: 65 of
US20160369298; herein SEQ ID NO: 1625), QEENNIAATPGVNT (SEQ ID NO:
66 of US20160369298; herein SEQ ID NO: 1626), QPPNNMAATHEVNT (SEQ
ID NO: 67 of US20160369298; herein SEQ ID NO: 1627), QHHNNSAATTIVNT
(SEQ ID NO: 68 of US20160369298; herein SEQ ID NO: 1628),
QTTNNRAAFNMVET (SEQ ID NO: 69 of US20160369298; herein SEQ ID NO:
1629), QKKNNNAASKKVAT (SEQ ID NO: 70 of US20160369298; herein SEQ
ID NO: 1630), QGGNNKAADDAVKT (SEQ ID NO: 71 of US20160369298;
herein SEQ ID NO: 1631), QAAKGGAADDAVKT (SEQ ID NO: 72 of
US20160369298; herein SEQ ID NO: 1632), QDDRAAAANESVDT (SEQ ID NO:
73 of US20160369298; herein SEQ ID NO: 1633), QQQHDDAAYQRVHT (SEQ
ID NO: 74 of US20160369298; herein SEQ ID NO: 1634), QSSSSLAAVSTVQT
(SEQ ID NO: 75 of US20160369298; herein SEQ ID NO: 1635),
QNNQTTAAIRNVTT (SEQ ID NO: 76 of US20160369298; herein SEQ ID NO:
1636), NYNKKSDNVDFT (SEQ ID NO: 77 of US20160369298; herein SEQ ID
NO: 1637), NYNKKSENVDFT (SEQ ID NO: 78 of US20160369298; herein SEQ
ID NO: 1638), NYNKKSLNVDFT (SEQ ID NO: 79 of US20160369298; herein
SEQ ID NO: 1639), NYNKKSPNVDFT (SEQ ID NO: 80 of US20160369298;
herein SEQ ID NO: 1640), NYSKKSHCVDFT (SEQ ID NO: 81 of
US20160369298; herein SEQ ID NO: 1641), NYRKTIYVDFT (SEQ ID NO: 82
of US20160369298; herein SEQ ID NO: 1642), NYKEKKDVHFT (SEQ ID NO:
83 of US20160369298; herein SEQ ID NO: 1643), NYGHRAIVQFT (SEQ ID
NO: 84 of US20160369298; herein SEQ ID NO: 1644), NYANHQFVVCT (SEQ
ID NO: 85 of US20160369298; herein SEQ ID NO: 1645), NYDDDPTGVLLT
(SEQ ID NO: 86 of US20160369298; herein SEQ ID NO: 1646),
NYDDPTGVLLT (SEQ ID NO: 87 of US20160369298; herein SEQ ID NO:
1647), NFEQQNSVEWT (SEQ ID NO: 88 of US20160369298; herein SEQ ID
NO: 1648), SQSGASN (SEQ ID NO: 89 and SEQ ID NO: 241 of
US20160369298; herein SEQ ID NO: 1649), NNGSQA (SEQ ID NO: 90 of
US20160369298; herein SEQ ID NO: 1650), YYLSRTNTPSGTTTWSRLQFSQAGA
(SEQ ID NO: 91 of US20160369298; herein SEQ ID NO: 1651),
SKTSADNNNSEYSWTG (SEQ ID NO: 92 of US20160369298; herein SEQ ID NO:
1652), HKDDEEKF (SEQ ID NO: 93, 209, 214, 219, 224, 234, 239, and
244 of US20160369298; herein SEQ ID NO: 1653), KQGSEKTNVDIEEV (SEQ
ID NO: 94 of US20160369298; herein SEQ ID NO: 1654), QRGNNQAATADVNT
(SEQ ID NO: 95 of US20160369298; herein SEQ ID NO: 1655),
NYNKKSVNVDFT (SEQ ID NO: 96 of US20160369298; herein SEQ ID NO:
1656), SQSGASNYNTPSGTTTQSRLQFSTSADNNNSEYSWTGATKYH (SEQ ID NO: 106
of US20160369298; herein SEQ ID NO: 1657),
SASGASNFNSEGGSLTQSSLGFSTDGENNNSDFSWTGATKYH (SEQ ID NO: 107 of
US20160369298; herein SEQ ID NO: 1658),
SQSGASNYNTPSGTTTQSRLQFSTDGENNNSDFSWTGATKYH (SEQ ID NO: 108 of
US20160369298; herein SEQ ID NO: 1659),
SASGASNYNTPSGTTTQSRLQFSTSADNNNSEFSWPGATTYH (SEQ ID NO: 109 of
US20160369298; herein SEQ ID NO: 1660),
SQSGASNFNSEGGSLTQSSLGFSTDGENNNSDFSWTGATKYH (SEQ ID NO: 110 of
US20160369298; herein SEQ ID NO: 1661),
SASGASNYNTPSGSLTQSSLGFSTDGENNNSDFSWTGATKYH (SEQ ID NO: 111 of
US20160369298; herein SEQ ID NO: 1662),
SQSGASNYNTPSGTTTQSRLQFSTSADNNNSDFSWTGATKYH (SEQ ID NO: 112 of
US20160369298; herein SEQ ID NO: 1663),
SGAGASNFNSEGGSLTQSSLGFSTDGENNNSDFSWTGATKYH (SEQ ID NO: 113 of
US20160369298; herein SEQ ID NO: 1664), SGAGASN (SEQ ID NO: 176 of
US20160369298; herein SEQ ID NO: 1665), NSEGGSLTQSSLGFS (SEQ ID NO:
177, 185, 193 and 202 of US20160369298; herein SEQ ID NO: 1666),
TDGENNNSDFS (SEQ ID NO: 178 of US20160369298; herein SEQ ID NO:
1667), SEFSWPGATT (SEQ ID NO: 179 of US20160369298; herein SEQ ID
NO: 1668), TSADNNNSDFSWT (SEQ ID NO: 180 of US20160369298; herein
SEQ ID NO: 1669), SQSGASNY (SEQ ID NO: 181, 187, and 198 of
US20160369298; herein SEQ ID NO: 1670), NTPSGTTTQSRLQFS (SEQ ID NO:
182, 188, 191, and 199 of US20160369298; herein SEQ ID NO: 1671),
TSADNNNSEYSWTGATKYH (SEQ ID NO: 183 of US20160369298; herein SEQ ID
NO: 1672), SASGASNF (SEQ ID NO: 184 of US20160369298; herein SEQ ID
NO: 1673), TDGENNNSDFSWTGATKYH (SEQ ID NO: 186, 189, 194, 197, and
203 of US20160369298; herein SEQ ID NO: 1674), SASGASNY (SEQ ID NO:
190 and SEQ ID NO: 195 of US20160369298; herein SEQ ID NO: 1675),
TSADNNNSEFSWPGATTYH (SEQ ID NO: 192 of US20160369298; herein SEQ ID
NO: 1676), NTPSGSLTQSSLGFS (SEQ ID NO: 196 of US20160369298; herein
SEQ ID NO: 1677), TSADNNNSDFSWTGATKYH (SEQ ID NO: 200 of
US20160369298; herein SEQ ID NO: 1678), SGAGASNF (SEQ ID NO: 201 of
US20160369298; herein SEQ ID NO: 1679),
CTCCAGVVSVVSMRSRVCVNSGCAGCTDHCVVSRNSGTCVMSACACAA (SEQ ID NO: 204 of
US20160369298; herein SEQ ID NO: 1680),
CTCCAGAGAGGCAACAGACAAGCAGCTACCGCAGATGTCAACACACAA (SEQ ID NO: 205 of
US20160369298; herein SEQ ID NO: 1681), SAAGASN (SEQ ID NO: 206 of
US20160369298; herein SEQ ID NO: 1682), YFLSRTNTESGSTTQSTLRFSQAG
(SEQ ID NO: 207 of US20160369298; herein SEQ ID NO: 1683),
SKTSADNNNSDFS (SEQ ID NO: 208, 228, and 253 of US20160369298;
herein SEQ ID NO: 1684), KQGSEKTDVDIDKV (SEQ ID NO: 210 of
US20160369298; herein SEQ ID NO: 1685), STAGASN (SEQ ID NO: 211 of
US20160369298; herein SEQ ID NO: 1686), YFLSRTNTTSGIETQSTLRFSQAG
(SEQ ID NO: 212 and SEQ ID NO: 247 of US20160369298; herein SEQ ID
NO: 1687), SKTDGENNNSDFS (SEQ ID NO: 213 and SEQ ID NO: 248 of
US20160369298; herein SEQ ID NO: 1688), KQGAAADDVEIDGV (SEQ ID NO:
215 and SEQ ID NO: 250 of US20160369298; herein SEQ ID NO: 1689),
SEAGASN (SEQ ID NO: 216 of US20160369298; herein SEQ ID NO: 1690),
YYLSRTNTPSGTTTQSRLQFSQAG (SEQ ID NO: 217, 232 and 242 of
US20160369298; herein SEQ ID NO: 1691), SKTSADNNNSEYS (SEQ ID NO:
218, 233, 238, and 243 of US20160369298; herein SEQ ID NO: 1692),
KQGSEKTNVDIEKV (SEQ ID NO: 220, 225 and 245 of US20160369298;
herein SEQ ID NO: 1693), YFLSRTNDASGSDTKSTLLFSQAG (SEQ ID NO: 222
of US20160369298; herein SEQ ID NO: 1694), STTPSENNNSEYS (SEQ ID
NO: 223 of US20160369298; herein SEQ ID NO: 1695), SAAGATN (SEQ ID
NO: 226 and SEQ ID NO: 251 of US20160369298; herein SEQ ID NO:
1696), YFLSRTNGEAGSATLSELRFSQAG (SEQ ID NO: 227 of US20160369298;
herein SEQ ID NO: 1697), HGDDADRF (SEQ ID NO: 229 and SEQ ID NO:
254 of US20160369298; herein SEQ ID NO: 1698), KQGAEKSDVEVDRV (SEQ
ID NO: 230 and SEQ ID NO: 255 of US20160369298; herein SEQ ID NO:
1699), KQDSGGDNIDIDQV (SEQ ID NO: 235 of US20160369298; herein SEQ
ID NO: 1700), SDAGASN (SEQ ID NO: 236 of US20160369298; herein SEQ
ID NO: 1701), YFLSRTNTEGGHDTQSTLRFSQAG (SEQ ID NO: 237 of
US20160369298; herein SEQ ID NO: 1702), KEDGGGSDVAIDEV (SEQ ID NO:
240 of US20160369298; herein SEQ ID NO: 1703), SNAGASN (SEQ ID NO:
246 of US20160369298; herein SEQ ID NO: 1704), and
YFLSRTNGEAGSATLSELRFSQPG (SEQ ID NO: 252 of US20160369298; herein
SEQ ID NO: 1705). Non-limiting examples of nucleotide sequences
that may encode the amino acid mutated sites include the following,
AGCVVMDCAGGARSCASCAAC (SEQ ID NO: 97 of US20160369298; herein SEQ
ID NO: 1706), AACRACRRSMRSMAGGCA (SEQ ID NO: 98 of US20160369298;
herein SEQ ID NO: 1707), CACRRGGACRRCRMSRRSARSTTT (SEQ ID NO: 99 of
US20160369298; herein SEQ ID NO: 1708),
TATTTCTTGAGCAGAACAAACRVCVVSRSCGGAMNCVHSACGMHSTCAVVSCTTVDSTT
TTCTCAGSBCRGSGCG (SEQ ID NO: 100 of US20160369298; herein SEQ ID
NO: 1709), TCAAMAMMAVNSRVCSRSAACAACAACAGTRASTTCTCGTGGMMAGGA (SEQ ID
NO: 101 of US20160369298; herein SEQ ID NO: 1710),
AAGSAARRCRSCRVSRVARVCRATRYCGMSNHCRVMVRSGTC (SEQ ID NO: 102 of
US20160369298; herein SEQ ID NO: 1711),
CAGVVSVVSMRSRVCVNSGCAGCTDHCVVSRNSGTCVMSACA (SEQ ID NO: 103 of
US20160369298; herein SEQ ID NO: 1712),
AACTWCRVSVASMVSVHSDDTGTGSWSTKSACT (SEQ ID NO: 104 of US20160369298;
herein SEQ ID NO: 1713), TTGTTGAACATCACCACGTGACGCACGTTC (SEQ ID NO:
256 of US20160369298; herein SEQ ID NO: 1714),
TCCCCGTGGTTCTACTACATAATGTGGCCG (SEQ ID NO: 257 of US20160369298;
herein SEQ ID NO: 1715), TTCCACACTCCGTTTTGGATAATGTTGAAC (SEQ ID NO:
258 of US20160369298; herein SEQ ID NO: 1716),
AGGGACATCCCCAGCTCCATGCTGTGGTCG (SEQ ID NO: 259 of US20160369298;
herein SEQ ID NO: 1717), AGGGACAACCCCTCCGACTCGCCCTAATCC (SEQ ID NO:
260 of US20160369298; herein SEQ ID NO: 1718),
TCCTAGTAGAAGACACCCTCTCACTGCCCG (SEQ ID NO: 261 of US20160369298;
herein SEQ ID NO: 1719), AGTACCATGTACACCCACTCTCCCAGTGCC (SEQ ID NO:
262 of US20160369298; herein SEQ ID NO: 1720),
ATATGGACGTTCATGCTGATCACCATACCG (SEQ ID NO: 263 of US20160369298;
herein SEQ ID NO: 1721), AGCAGGAGCTCCTTGGCCTCAGCGTGCGAG (SEQ ID NO:
264 of US20160369298; herein SEQ ID NO: 1722),
ACAAGCAGCTTCACTATGACAACCACTGAC (SEQ ID NO: 265 of US20160369298;
herein SEQ ID NO: 1723),
CAGCCTAGGAACTGGCTTCCTGGACCCTGTTACCGCCAGCAGAGAGTCTCAAMAMMAV
NSRVCSRSAACAACAACAGTRASTTCTCCTGGMMAGGAGCTACCAAGTACCACCTCAAT
GGCAGAGACTCTCTGGTGAATCCCGGACCAGCTATGGCAAGCCACRRGGACRRCRMSR
RSARSTTTTTTCCTCAGAGCGGGGTTCTCATCTTTGGGAAGSAARRCRSCRVSRVARVCR
ATRYCGMSNHCRVMVRSGTCATGATTACAGACGAAGAGGAGATCTGGAC (SEQ ID NO: 266
of US20160369298; herein SEQ ID NO: 1724),
TGGGACAATGGCGGTCGTCTCTCAGAGTTKTKKT (SEQ ID NO: 267 of
US20160369298; herein SEQ ID NO: 1725),
AGAGGACCKKTCCTCGATGGTTCATGGTGGAGTTA (SEQ ID NO: 268 of
US20160369298; herein SEQ ID NO: 1726),
CCACTTAGGGCCTGGTCGATACCGTTCGGTG (SEQ ID NO: 269 of US20160369298;
herein SEQ ID NO: 1727), and TCTCGCCCCAAGAGTAGAAACCCTTCSTTYYG (SEQ
ID NO: 270 of US20160369298; herein SEQ ID NO: 1728).
[0075] In some embodiments, the AAV serotype may comprise an ocular
cell targeting peptide as described in International Patent
Publication WO2016134375, the contents of which are herein
incorporated by reference in their entirety, such as, but not
limited to SEQ ID NO: 9, and SEQ ID NO:10 of WO2016134375. Further,
any of the ocular cell targeting peptides or amino acids described
in WO2016134375, may be inserted into any parent AAV serotype, such
as, but not limited to, AAV2 (SEQ ID NO:8 of WO2016134375; herein
SEQ ID NO: 1729), or AAV9 (SEQ ID NO: 11 of WO2016134375; herein
SEQ ID NO: 1730). In some embodiments, modifications, such as
insertions are made in AAV2 proteins at P34-A35, T138-A139,
A139-P140, G453-T454, N587-R588, and/or R588-Q589. In certain
embodiments, insertions are made at D384, G385, 1560, T561, N562,
E563, E564, E565, N704, and/or Y705 of AAV9. The ocular cell
targeting peptide may be, but is not limited to, any of the
following amino acid sequences, GSTPPPM (SEQ ID NO: 1 of
WO2016134375; herein SEQ ID NO: 1731), or GETRAPL (SEQ ID NO: 4 of
WO2016134375; herein SEQ ID NO: 1732).
[0076] In some embodiments, the AAV serotype may be modified as
described in the United States Publication US 20170145405 the
contents of which are herein incorporated by reference in their
entirety. AAV serotypes may include, modified AAV2 (e.g.,
modifications at Y444F, Y500F, Y730F and/or S662V), modified AAV3
(e.g., modifications at Y705F, Y731F and/or T492V), and modified
AAV6 (e.g., modifications at S663V and/or T492V).
[0077] In some embodiments, the AAV serotype may be modified as
described in the International Publication WO2017083722 the
contents of which are herein incorporated by reference in their
entirety. AAV serotypes may include, AAV1 (Y705+731F+T492V), AAV2
(Y444+500+730F+T491V), AAV3 (Y705+731F), AAV5, AAV 5
(Y436+693+719F), AAV6 (VP3 variant Y705F/Y731F/T492V), AAV8
(Y733F), AAV9, AAV9 (VP3 variant Y731F), and AAV10 (Y733F).
[0078] In some embodiments, the AAV serotype may comprise, as
described in International Patent Publication WO2017015102, the
contents of which are herein incorporated by reference in their
entirety, an engineered epitope comprising the amino acids SPAKFA
(SEQ ID NO: 24 of WO2017015102; herein SEQ ID NO: 1733) or NKDKLN
(SEQ ID NO:2 of WO2017015102; herein SEQ ID NO: 1734). The epitope
may be inserted in the region of amino acids 665 to 670 based on
the numbering of the VP1 capsid of AAV8 (SEQ ID NO:3 of
WO2017015102) and/or residues 664 to 668 of AAV3B (SEQ ID
NO:3).
[0079] In one embodiment, the AAV serotype may be, or may have a
sequence as described in International Patent Publication
WO2017058892, the contents of which are herein incorporated by
reference in their entirety, such as, but not limited to, AAV
variants with capsid proteins that may comprise a substitution at
one or more (e.g., 2, 3, 4, 5, 6, or 7) of amino acid residues
262-268, 370-379, 451-459, 472-473, 493-500, 528-534, 547-552,
588-597, 709-710, 716-722 of AAV1, in any combination, or the
equivalent amino acid residues in AAV2, AAV3, AAV4, AAV5, AAV6,
AAV7, AAV8, AAV9, AAV10, AAV11, AAV12, AAVrh8, AAVrh10, AAVrh32.33,
bovine AAV or avian AAV. The amino acid substitution may be, but is
not limited to, any of the amino acid sequences described in
WO2017058892. In one embodiment, the AAV may comprise an amino acid
substitution at residues 256L, 258K, 259Q, 261S, 263A, 264S, 265T,
266G, 272H, 385S, 386Q, S472R, V473D, N500E 547S, 709A, 710N, 716D,
717N, 718N, 720L, A456T, Q457T, N458Q, K459S, T492S, K493A, S586R,
S587G, S588N, T589R and/or 722T of AAV1 (SEQ ID NO: 1 of
WO2017058892) in any combination, 244N, 246Q, 248R, 249E, 2501,
251K, 252S, 253G, 254S, 255V, 256D, 263Y, 377E, 378N, 453L, 456R,
532Q, 533P, 535N, 536P, 537G, 538T, 539T, 540A, 541T, 542Y, 543L,
546N, 653V, 654P, 656S, 697Q, 698F, 704D, 705S, 706T, 707G, 708E,
709Y and/or 710R of AAV5 (SEQ ID NO:5 of WO2017058892) in any
combination, 248R, 316V, 317Q, 318D, 319S, 443N, 530N, 531S, 532Q
533P, 534A, 535N, 540A, 541 T, 542Y, 543L, 545G, 546N, 697Q, 704D,
706T, 708E, 709Y and/or 710R of AAV5 (SEQ ID NO: 5 of WO2017058892)
in any combination, 264S, 266G, 269N, 272H, 457Q, 588S and/or 5891
of AAV6 (SEQ ID NO:6 WO2017058892) in any combination, 457T, 459N,
496G, 499N, 500N, 589Q, 590N and/or 592A of AAV8 (SEQ ID NO: 8
WO2017058892) in any combination, 451I, 452N, 453G, 454S, 455G,
456Q, 457N and/or 458Q of AAV9 (SEQ ID NO: 9 WO2017058892) in any
combination.
[0080] In some embodiments, the AAV may include a sequence of amino
acids at positions 155, 156 and 157 of VP1 or at positions 17, 18,
19 and 20 of VP2, as described in International Publication No. WO
2017066764, the contents of which are herein incorporated by
reference in their entirety. The sequences of amino acid may be,
but not limited to, N-S-S, S-X-S, S-S-Y, N-X-S, N-S-Y, S-X-Y and
N-X-Y, where N, X and Y are, but not limited to, independently
non-serine, or non-threonine amino acids, wherein the AAV may be,
but not limited to AAV1, AAV2, AAV3, AAV4, AAV5, AAV6, AAV7, AAV8,
AAV9, AAV10, AAV11 and AAV12. In some embodiments, the AAV may
include a deletion of at least one amino acid at positions 156, 157
or 158 of VP1 or at positions 19, 20 or 21 of VP2, wherein the AAV
may be, but not limited to AAV1, AAV2, AAV3, AAV4, AAV5, AAV6,
AAV7, AAV8, AAV9, AAV10, AAV11 and AAV12.
Viral Genome Component: Inverted Terminal Repeats (ITRs)
[0081] The AAV particles of the present disclosure comprise a viral
genome with at least one ITR region and a payload region. In one
embodiment, the viral genome has two ITRs. These two ITRs flank the
payload region at the 5' and 3' ends. The ITRs function as origins
of replication comprising recognition sites for replication. ITRs
comprise sequence regions which can be complementary and
symmetrically arranged. ITRs incorporated into viral genomes of the
disclosure may be comprised of naturally occurring polynucleotide
sequences or recombinantly derived polynucleotide sequences.
[0082] The ITRs may be derived from the same serotype as the
capsid, selected from any of the serotypes listed in Table 1, or a
derivative thereof. The ITR may be of a different serotype than the
capsid. In one embodiment, the AAV particle has more than one ITR.
In a non-limiting example, the AAV particle has a viral genome
comprising two ITRs. In one embodiment, the ITRs are of the same
serotype as one another. In another embodiment, the ITRs are of
different serotypes. Non-limiting examples include zero, one or
both of the ITRs having the same serotype as the capsid. In one
embodiment both ITRs of the viral genome of the AAV particle are
AAV2 ITRs.
[0083] Independently, each ITR may be about 100 to about 150
nucleotides in length. An ITR may be about 100-105 nucleotides in
length, 106-110 nucleotides in length, 111-115 nucleotides in
length, 116-120 nucleotides in length, 121-125 nucleotides in
length, 126-130 nucleotides in length, 131-135 nucleotides in
length, 136-140 nucleotides in length, 141-145 nucleotides in
length or 146-150 nucleotides in length. In one embodiment, the
ITRs are 140-142 nucleotides in length. Non-limiting examples of
ITR length are 102, 140, 141, 142, 145 nucleotides in length, and
those having at least 95% identity thereto.
Viral Genome Component: Promoters
[0084] In one embodiment, the payload region of the viral genome
comprises at least one element to enhance the transgene target
specificity and expression (See e.g., Powell et al. Viral
Expression Cassette Elements to Enhance Transgene Target
Specificity and Expression in Gene Therapy, 2015; the contents of
which are herein incorporated by reference in its entirety).
Non-limiting examples of elements to enhance the transgene target
specificity and expression include promoters, endogenous miRNAs,
post-transcriptional regulatory elements (PREs), polyadenylation
(PolyA) signal sequences and upstream enhancers (USEs), CMV
enhancers and introns.
[0085] A person skilled in the art may recognize that expression of
the polypeptides of the disclosure in a target cell may require a
specific promoter, including but not limited to, a promoter that is
species specific, inducible, tissue-specific, or cell
cycle-specific (Parr et al., Nat. Med. 3:1145-9 (1997); the
contents of which are herein incorporated by reference in their
entirety).
[0086] In one embodiment, the promoter is deemed to be efficient
when it drives expression of the polypeptide(s) encoded in the
payload region of the viral genome of the AAV particle. As
anon-limiting example, that polypeptide is AADC.
[0087] In one embodiment, the promoter is a promoter deemed to be
efficient when it drives expression in the cell being targeted.
[0088] In one embodiment, the promoter is a promoter having a
tropism for the cell being targeted.
[0089] In one embodiment, the promoter drives expression of the
payload for a period of time in targeted tissues. Expression driven
by a promoter may be for a period of 1 hour, 2, hours, 3 hours, 4
hours, 5 hours, 6 hours, 7 hours, 8 hours, 9 hours, 10 hours, 11
hours, 12 hours, 13 hours, 14 hours, 15 hours, 16 hours, 17 hours,
18 hours, 19 hours, 20 hours, 21 hours, 22 hours, 23 hours, 1 day,
2 days, 3 days, 4 days, 5 days, 6 days, 1 week, 8 days, 9 days, 10
days, 11 days, 12 days, 13 days, 2 weeks, 15 days, 16 days, 17
days, 18 days, 19 days, 20 days, 3 weeks, 22 days, 23 days, 24
days, 25 days, 26 days, 27 days, 28 days, 29 days, 30 days, 31
days, 1 month, 2 months, 3 months, 4 months, 5 months, 6 months, 7
months, 8 months, 9 months, 10 months, 11 months, 1 year, 13
months, 14 months, 15 months, 16 months, 17 months, 18 months, 19
months, 20 months, 21 months, 22 months, 23 months, 2 years, 3
years, 4 years, 5 years, 6 years, 7 years, 8 years, 9 years, 10
years or more than 10 years. Expression may be for 1-5 hours, 1-12
hours, 1-2 days, 1-5 days, 1-2 weeks, 1-3 weeks, 1-4 weeks, 1-2
months, 1-4 months, 1-6 months, 2-6 months, 3-6 months, 3-9 months,
4-8 months, 6-12 months, 1-2 years, 1-5 years, 2-5 years, 3-6
years, 3-8 years, 4-8 years or 5-10 years. As a non-limiting
example, the promoter is a weak promoter for sustained expression
of a payload in nervous tissues.
[0090] In one embodiment, the promoter drives expression of the
polypeptides of the disclosure for at least 1 month, 2 months, 3
months, 4 months, 5 months, 6 months, 7 months, 8 months, 9 months,
10 months, 11 months, 1 year, 2 years, 3 years 4 years, 5 years, 6
years, 7 years, 8 years, 9 years, 10 years, 11 years, 12 years, 13
years, 14 years, 15 years, 16 years, 17 years, 18 years, 19 years,
20 years, 21 years, 22 years, 23 years, 24 years, 25 years, 26
years, 27 years, 28 years, 29 years, 30 years, 31 years, 32 years,
33 years, 34 years, 35 years, 36 years, 37 years, 38 years, 39
years, 40 years, 41 years, 42 years, 43 years, 44 years, 45 years,
46 years, 47 years, 48 years, 49 years, 50 years, 55 years, 60
years, 65 years, or more than 65 years.
[0091] Promoters may be naturally occurring or non-naturally
occurring. Non-limiting examples of promoters include viral
promoters, plant promoters and mammalian promoters. In some
embodiments, the promoters may be human promoters. In some
embodiments, the promoter may be truncated.
[0092] Promoters which drive or promote expression in most tissues
include, but are not limited to, human elongation factor
1.alpha.-subunit (EF1.alpha.), cytomegalovirus (CMV)
immediate-early enhancer and/or promoter, chicken .beta.-actin
(CBA) and its derivative CAG, .beta. glucuronidase (GUSB), or
ubiquitin C (UBC). Tissue-specific expression elements can be used
to restrict expression to certain cell types such as, but not
limited to, muscle specific promoters, B cell promoters, monocyte
promoters, leukocyte promoters, macrophage promoters, pancreatic
acinar cell promoters, endothelial cell promoters, lung tissue
promoters, astrocyte promoters, or nervous system promoters which
can be used to restrict expression to neurons, astrocytes, or
oligodendrocytes.
[0093] Non-limiting examples of muscle-specific promoters include
mammalian muscle creatine kinase (MCK) promoter, mammalian desmin
(DES) promoter, mammalian troponin I (TNNI2) promoter, and
mammalian skeletal alpha-actin (ASKA) promoter (see, e.g. U.S.
Patent Publication US 20110212529, the contents of which are herein
incorporated by reference in their entirety).
[0094] Non-limiting examples of tissue-specific expression elements
for neurons include neuron-specific enolase (NSE), platelet-derived
growth factor (PDGF), platelet-derived growth factor B-chain
(PDGF-.beta.), synapsin (Syn), methyl-CpG binding protein 2
(MeCP2), Ca.sup.2+/calmodulin-dependent protein kinase II (CaMKII),
metabotropic glutamate receptor 2 (mGluR2), neurofilament light
(NFL) or heavy (NFH), .beta.-globin minigene np32, preproenkephalin
(PPE), enkephalin (Enk) and excitatory amino acid transporter 2
(EAAT2) promoters. Non-limiting examples of tissue-specific
expression elements for astrocytes include glial fibrillary acidic
protein (GFAP) and EAAT2 promoters. A non-limiting example of a
tissue-specific expression element for oligodendrocytes includes
the myelin basic protein (MBP) promoter.
[0095] In one embodiment, the promoter may be less than 1 kb. The
promoter may have a length of 200, 210, 220, 230, 240, 250, 260,
270, 280, 290, 300, 310, 320, 330, 340, 350, 360, 370, 380, 390,
400, 410, 420, 430, 440, 450, 460, 470, 480, 490, 500, 510, 520,
530, 540, 550, 560, 570, 580, 590, 600, 610, 620, 630, 640, 650,
660, 670, 680, 690, 700, 710, 720, 730, 740, 750, 760, 770, 780,
790, 800 or more than 800 nucleotides. The promoter may have a
length between 200-300, 200-400, 200-500, 200-600, 200-700,
200-800, 300-400, 300-500, 300-600, 300-700, 300-800, 400-500,
400-600, 400-700, 400-800, 500-600, 500-700, 500-800, 600-700,
600-800 or 700-800.
[0096] In one embodiment, the promoter may be a combination of two
or more components of the same or different starting or parental
promoters such as, but not limited to, CMV and CBA. Each component
may have a length of 200, 210, 220, 230, 240, 250, 260, 270, 280,
290, 300, 310, 320, 330, 340, 350, 360, 370, 380, 381, 382, 383,
384, 385, 386, 387, 388, 389, 390, 400, 410, 420, 430, 440, 450,
460, 470, 480, 490, 500, 510, 520, 530, 540, 550, 560, 570, 580,
590, 600, 610, 620, 630, 640, 650, 660, 670, 680, 690, 700, 710,
720, 730, 740, 750, 760, 770, 780, 790, 800 or more than 800. Each
component may have a length between 200-300, 200-400, 200-500,
200-600, 200-700, 200-800, 300-400, 300-500, 300-600, 300-700,
300-800, 400-500, 400-600, 400-700, 400-800, 500-600, 500-700,
500-800, 600-700, 600-800 or 700-800. In one embodiment, the
promoter is a combination of a 382 nucleotide CMV-enhancer sequence
and a 260 nucleotide CBA-promoter sequence.
[0097] In one embodiment, the viral genome comprises a ubiquitous
promoter. Non-limiting examples of ubiquitous promoters include
CMV, CBA (including derivatives CAG, CBh, etc.), EF-1.alpha., PGK,
UBC, GUSB (hGBp), and UCOE (promoter of HNRPA2B1-CBX3).
[0098] Yu et al. (Molecular Pain 2011, 7:63; the contents of which
are herein incorporated by reference in their entirety) evaluated
the expression of eGFP under the CAG, EFI.alpha., PGK and UBC
promoters in rat DRG cells and primary DRG cells using lentiviral
vectors and found that UBC showed weaker expression than the other
3 promoters and only 10-12% glial expression was seen for all
promoters. Soderblom et al. (E. Neuro 2015; the contents of which
are herein incorporated by reference in its entirety) evaluated the
expression of eGFP in AAV8 with CMV and UBC promoters and AAV2 with
the CMV promoter after injection in the motor cortex. Intranasal
administration of a plasmid containing a UBC or EFIa promoter
showed a sustained airway expression greater than the expression
with the CMV promoter (See e.g., Gill et al., Gene Therapy 2001,
Vol. 8, 1539-1546; the contents of which are herein incorporated by
reference in their entirety). Husain et al. (Gene Therapy 2009; the
contents of which are herein incorporated by reference in its
entirety) evaluated an H.beta.H construct with a hGUSB promoter, a
HSV-1LAT promoter and an NSE promoter and found that the H.beta.H
construct showed weaker expression than NSE in mouse brain. Passini
and Wolfe (J. Virol. 2001, 12382-12392, the contents of which are
herein incorporated by reference in its entirety) evaluated the
long term effects of the H.beta.H vector following an
intraventricular injection in neonatal mice and found that there
was sustained expression for at least 1 year. Low expression in all
brain regions was found by Xu et al. (Gene Therapy 2001, 8,
1323-1332; the contents of which are herein incorporated by
reference in their entirety) when NFL and NFH promoters were used
as compared to the CMV-lacZ, CMV-luc, EF, GFAP, hENK, nAChR, PPE,
PPE+wpre, NSE (0.3 kb), NSE (1.8 kb) and NSE (1.8 kb+wpre). Xu et
al. found that the promoter activity in descending order was NSE
(1.8 kb), EF, NSE (0.3 kb), GFAP, CMV, hENK, PPE, NFL and NFH. NFL
is a 650 nucleotide promoter and NFH is a 920 nucleotide promoter
which are both absent in the liver but NFH is abundant in the
sensory proprioceptive neurons, brain and spinal cord and NFH is
present in the heart. SCN8A is a 470 nucleotide promoter which
expresses throughout the DRG, spinal cord and brain with
particularly high expression seen in the hippocampal neurons and
cerebellar Purkinje cells, cortex, thalamus and hypothalamus (See
e.g., Drews et al. Identification of evolutionary conserved,
functional noncoding elements in the promoter region of the sodium
channel gene SCN8A, Mamm Genome (2007) 18:723-731; and Raymond et
al. Expression of Alternatively Spliced Sodium Channel a-subunit
genes, Journal of Biological Chemistry (2004) 279(44) 46234-46241;
the contents of each of which are herein incorporated by reference
in their entireties).
[0099] Any of promoters taught by the aforementioned Yu, Soderblom,
Gill, Husain, Passini, Xu, Drews or Raymond may be used in the
present disclosures.
[0100] In one embodiment, the promoter is not cell specific.
[0101] In one embodiment, the promoter is an ubiquitin c (UBC)
promoter. The UBC promoter may have a size of 300-350 nucleotides.
As a non-limiting example, the UBC promoter is 332 nucleotides.
[0102] In one embodiment, the promoter is a .beta.-glucuronidase
(GUSB) promoter. The GUSB promoter may have a size of 350-400
nucleotides. As a non-limiting example, the GUSB promoter is 378
nucleotides.
[0103] In one embodiment, the promoter is a neurofilament light
(NFL) promoter. The NFL promoter may have a size of 600-700
nucleotides. As a non-limiting example, the NFL promoter is 650
nucleotides.
[0104] In one embodiment, the promoter is a neurofilament heavy
(NFH) promoter. The NFH promoter may have a size of 900-950
nucleotides. As a non-limiting example, the NFH promoter is 920
nucleotides.
[0105] In one embodiment, the promoter is a SCN8A promoter. The
SCN8A promoter may have a size of 450-500 nucleotides. As a
non-limiting example, the SCN8A promoter is 470 nucleotides.
[0106] In one embodiment, the promoter is a frataxin (FXN)
promoter. The FXN promoter may also be referred to as the FRDA
promoter.
[0107] In one embodiment, the promoter is a phosphoglycerate kinase
1 (PGK) promoter.
[0108] In one embodiment, the promoter is a chicken .beta.-actin
(CBA) promoter.
[0109] In one embodiment, the promoter is a cytomegalovirus (CMV)
promoter.
[0110] In one embodiment, the promoter is a H1 promoter.
[0111] In one embodiment, the promoter is an engineered
promoter.
[0112] In one embodiment, the promoter is a liver or a skeletal
muscle promoter. Non-limiting examples of liver promoters include
human .alpha.-1-antitrypsin (hAAT) and thyroxine binding globulin
(TBG). Non-limiting examples of skeletal muscle promoters include
Desmin, MCK or synthetic C5-12.
[0113] In one embodiment, the promoter is a RNA pol III promoter.
As a non-limiting example, the RNA pol III promoter is U6. As a
non-limiting example, the RNA pol III promoter is H1.
[0114] In one embodiment, the viral genome comprises two promoters.
As a non-limiting example, the promoters are an EF1a promoter and a
CMV promoter.
[0115] In one embodiment, the viral genome comprises an enhancer
element, a promoter and/or a 5'UTR intron. The enhancer element,
also referred to herein as an "enhancer," may be, but is not
limited to, a CMV enhancer, the promoter may be, but is not limited
to, a CMV, CBA, UBC, GUSB, NSE, Synapsin, MeCP2, and GFAP promoter
and the 5'UTR/intron may be, but is not limited to, SV40, and
CBA-MVM. As a non-limiting example, the enhancer, promoter and/or
intron used in combination may be: (1) CMV enhancer, CMV promoter,
SV40 5'UTR intron; (2) CMV enhancer, CBA promoter, SV 40 5'UTR
intron; (3) CMV enhancer, CBA promoter, CBA-MVM 5'UTR intron; (4)
UBC promoter; (5) GUSB promoter; (6) NSE promoter; (7) Synapsin
promoter; (8) MeCP2 promoter and (9) GFAP promoter.
[0116] In one embodiment, the viral genome comprises an engineered
promoter.
[0117] In another embodiment, the viral genome comprises a promoter
from a naturally expressed protein.
[0118] In one embodiment, a region located approximately .about.5
kb upstream of the first exon of the payload in order to allow for
expression of the payload with the promoter. (See e.g., Puspasari
et al. Long Range Regulation of Human FXN Gene Expression, PLOS
ONE, 2011; the contents of which is herein incorporated by
reference in its entirety; a 17 bp region located approximately 4.9
kb upstream of the first exon of the frataxin gene in order to
allow for expression with the FRDA promoter).
[0119] In one embodiment, the vector genome may comprise a promoter
such as, but not limited to, CMV or U6. As a non-limiting example,
the promoter for the AAV particles comprising the payload of the
present disclosure is a CMV promoter. As another non-limiting
example, the promoter for the AAV particles comprising the payload
of the present disclosure is a U6 promoter.
[0120] In one embodiment, the vector genome may comprise a CMV and
a U6 promoter.
[0121] In one embodiment, the vector genome may comprise a CBA
promoter.
Viral Genome Component: Untranslated Regions (UTRs)
[0122] By definition, wild type untranslated regions (UTRs) of a
gene are transcribed but not translated. Generally, the 5' UTR
starts at the transcription start site and ends at the start codon
and the 3' UTR starts immediately following the stop codon and
continues until the termination signal for transcription.
[0123] Features typically found in abundantly expressed genes of
specific target organs may be engineered into UTRs to enhance the
stability and protein production. As a non-limiting example, a 5'
UTR from mRNA normally expressed in the liver (e.g., albumin, serum
amyloid A, Apolipoprotein A/B/E, transferrin, alpha fetoprotein,
erythropoietin, or Factor VIII) may be used in the viral genomes of
the AAV particles of the disclosure to enhance expression in
hepatic cell lines or liver.
[0124] While not wishing to be bound by theory, wild-type 5'
untranslated regions (UTRs) include features which play roles in
translation initiation. Kozak sequences, which are commonly known
to be involved in the process by which the ribosome initiates
translation of many genes, are usually included in 5' UTRs. Kozak
sequences have the consensus CCR(A/G)CCAUGG, where R is a purine
(adenine or guanine) three bases upstream of the start codon (ATG),
which is followed by another `G`.
[0125] In one embodiment, the 5'UTR in the viral genome includes a
Kozak sequence.
[0126] In one embodiment, the 5'UTR in the viral genome does not
include a Kozak sequence.
[0127] While not wishing to be bound by theory, wild-type 3' UTRs
are known to have stretches of Adenosines and Uridines embedded
therein. These AU rich signatures are particularly prevalent in
genes with high rates of turnover. Based on their sequence features
and functional properties, the AU rich elements (AREs) can be
separated into three classes (Chen et al, 1995, the contents of
which are herein incorporated by reference in its entirety): Class
I AREs, such as, but not limited to, c-Myc and MyoD, contain
several dispersed copies of an AUUUA motif within U-rich regions.
Class II AREs, such as, but not limited to, GM-CSF and TNF-a,
possess two or more overlapping UUAUUUA(U/A)(U/A) nonamers. Class
III ARES, such as, but not limited to, c-Jun and Myogenin, are less
well defined. These U rich regions do not contain an AUUUA motif.
Most proteins binding to the AREs are known to destabilize the
messenger, whereas members of the ELAV family, most notably HuR,
have been documented to increase the stability of mRNA. HuR binds
to AREs of all the three classes. Engineering the HuR specific
binding sites into the 3' UTR of nucleic acid molecules will lead
to HuR binding and thus, stabilization of the message in vivo.
[0128] Introduction, removal or modification of 3' UTR AU rich
elements (AREs) can be used to modulate the stability of
polynucleotides. When engineering specific polynucleotides, e.g.,
payload regions of viral genomes, one or more copies of an ARE can
be introduced to make polynucleotides less stable and thereby
curtail translation and decrease production of the resultant
protein. Likewise, AREs can be identified and removed or mutated to
increase the intracellular stability and thus increase translation
and production of the resultant protein.
[0129] In one embodiment, the 3' UTR of the viral genome may
include an oligo(dT) sequence for templated addition of a poly-A
tail.
[0130] In one embodiment, the viral genome may include at least one
miRNA seed, binding site or full sequence. microRNAs (or miRNA or
miR) are 19-25 nucleotide noncoding RNAs that bind to the sites of
nucleic acid targets and down-regulate gene expression either by
reducing nucleic acid molecule stability or by inhibiting
translation. A microRNA sequence comprises a "seed" region, i.e., a
sequence in the region of positions 2-8 of the mature microRNA,
which sequence has perfect Watson-Crick complementarity to the
miRNA target sequence of the nucleic acid.
[0131] In one embodiment, the viral genome may be engineered to
include, alter or remove at least one miRNA binding site, sequence
or seed region.
[0132] Any UTR from any gene known in the art may be incorporated
into the viral genome of the AAV particle. These UTRs, or portions
thereof, may be placed in the same orientation as in the gene from
which they were selected or they may be altered in orientation or
location. In one embodiment, the UTR used in the viral genome of
the AAV particle may be inverted, shortened, lengthened, made with
one or more other 5' UTRs or 3' UTRs known in the art. As used
herein, the term "altered" as it relates to a UTR, means that the
UTR has been changed in some way in relation to a reference
sequence. For example, a 3' or 5' UTR may be altered relative to a
wild type or native UTR by the change in orientation or location as
taught above or may be altered by the inclusion of additional
nucleotides, deletion of nucleotides, swapping or transposition of
nucleotides.
[0133] In one embodiment, the viral genome of the AAV particle
comprises at least one artificial UTRs which is not a variant of a
wild type UTR.
[0134] In one embodiment, the viral genome of the AAV particle
comprises UTRs which have been selected from a family of
transcripts whose proteins share a common function, structure,
feature or property.
Viral Genome Component: Polyadenylation Sequence
[0135] In one embodiment, the viral genome of the AAV particles of
the present disclosure comprise at least one polyadenylation
sequence. The viral genome of the AAV particle may comprise a
polyadenylation sequence between the 3' end of the payload coding
sequence and the 5' end of the 3'ITR.
[0136] In one embodiment, the polyadenylation sequence or "polyA
sequence" may range from absent to about 500 nucleotides in length.
The polyadenylation sequence may be, but is not limited to, 1, 2,
3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20,
21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37,
38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54,
55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71,
72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88,
89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103,
104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116,
117, 118, 119, 120, 121, 122, 123, 124, 125, 126, 127, 128, 129,
130, 131, 132, 133, 134, 135, 136, 137, 138, 139, 140, 141, 142,
143, 144, 145, 146, 147, 148, 149, 150, 151, 152, 153, 154, 155,
156, 157, 158, 159, 160, 161, 162, 163, 164, 165, 166, 167, 168,
169, 170, 171, 172, 173, 174, 175, 176, 177, 178, 179, 180, 181,
182, 183, 184, 185, 186, 187, 188, 189, 190, 191, 192, 193, 194,
195, 196, 197, 198, 199, 200, 201, 202, 203, 204, 205, 206, 207,
208, 209, 210, 211, 212, 213, 214, 215, 216, 217, 218, 219, 220,
221, 222, 223, 224, 225, 226, 227, 228, 229, 230, 231, 232, 233,
234, 235, 236, 237, 238, 239, 240, 241, 242, 243, 244, 245, 246,
247, 248, 249, 250, 251, 252, 253, 254, 255, 256, 257, 258, 259,
260, 261, 262, 263, 264, 265, 266, 267, 268, 269, 270, 271, 272,
273, 274, 275, 276, 277, 278, 279, 280, 281, 282, 283, 284, 285,
286, 287, 288, 289, 290, 291, 292, 293, 294, 295, 296, 297, 298,
299, 300, 301, 302, 303, 304, 305, 306, 307, 308, 309, 310, 311,
312, 313, 314, 315, 316, 317, 318, 319, 320, 321, 322, 323, 324,
325, 326, 327, 328, 329, 330, 331, 332, 333, 334, 335, 336, 337,
338, 339, 340, 341, 342, 343, 344, 345, 346, 347, 348, 349, 350,
351, 352, 353, 354, 355, 356, 357, 358, 359, 360, 361, 362, 363,
364, 365, 366, 367, 368, 369, 370, 371, 372, 373, 374, 375, 376,
377, 378, 379, 380, 381, 382, 383, 384, 385, 386, 387, 388, 389,
390, 391, 392, 393, 394, 395, 396, 397, 398, 399, 400, 401, 402,
403, 404, 405, 406, 407, 408, 409, 410, 411, 412, 413, 414, 415,
416, 417, 418, 419, 420, 421, 422, 423, 424, 425, 426, 427, 428,
429, 430, 431, 432, 433, 434, 435, 436, 437, 438, 439, 440, 441,
442, 443, 444, 445, 446, 447, 448, 449, 450, 451, 452, 453, 454,
455, 456, 457, 458, 459, 460, 461, 462, 463, 464, 465, 466, 467,
468, 469, 470, 471, 472, 473, 474, 475, 476, 477, 478, 479, 480,
481, 482, 483, 484, 485, 486, 487, 488, 489, 490, 491, 492, 493,
494, 495, 496, 497, 498, 499, and 500 nucleotides in length.
[0137] In one embodiment, the polyadenylation sequence is 50-100
nucleotides in length.
[0138] In one embodiment, the polyadenylation sequence is 50-150
nucleotides in length.
[0139] In one embodiment, the polyadenylation sequence is 50-160
nucleotides in length.
[0140] In one embodiment, the polyadenylation sequence is 50-200
nucleotides in length.
[0141] In one embodiment, the polyadenylation sequence is 60-100
nucleotides in length.
[0142] In one embodiment, the polyadenylation sequence is 60-150
nucleotides in length.
[0143] In one embodiment, the polyadenylation sequence is 60-160
nucleotides in length.
[0144] In one embodiment, the polyadenylation sequence is 60-200
nucleotides in length.
[0145] In one embodiment, the polyadenylation sequence is 70-100
nucleotides in length.
[0146] In one embodiment, the polyadenylation sequence is 70-150
nucleotides in length.
[0147] In one embodiment, the polyadenylation sequence is 70-160
nucleotides in length.
[0148] In one embodiment, the polyadenylation sequence is 70-200
nucleotides in length.
[0149] In one embodiment, the polyadenylation sequence is 80-100
nucleotides in length.
[0150] In one embodiment, the polyadenylation sequence is 80-150
nucleotides in length.
[0151] In one embodiment, the polyadenylation sequence is 80-160
nucleotides in length.
[0152] In one embodiment, the polyadenylation sequence is 80-200
nucleotides in length.
[0153] In one embodiment, the polyadenylation sequence is 90-100
nucleotides in length.
[0154] In one embodiment, the polyadenylation sequence is 90-150
nucleotides in length.
[0155] In one embodiment, the polyadenylation sequence is 90-160
nucleotides in length.
[0156] In one embodiment, the polyadenylation sequence is 90-200
nucleotides in length.
Viral Genome Component: Introns
[0157] In one embodiment, the payload region comprises at least one
element to enhance the expression such as one or more introns or
portions thereof. Non-limiting examples of introns include, MVM
(67-97 bps), F.IX truncated intron 1 (300 bps), .beta.-globin
SD/immunoglobulin heavy chain splice acceptor (250 bps), adenovirus
splice donor/immunoglobin splice acceptor (500 bps), SV40 late
splice donor/splice acceptor (19S/16S) (180 bps) and hybrid
adenovirus splice donor/IgG splice acceptor (230 bps).
[0158] In one embodiment, the intron or intron portion may be
100-500 nucleotides in length. The intron may have a length of 80,
90, 100, 110, 120, 130, 140, 150, 160, 170, 171, 172, 173, 174,
175, 176, 177, 178, 179, 180, 190, 200, 210, 220, 230, 240, 250,
260, 270, 280, 290, 300, 310, 320, 330, 340, 350, 360, 370, 380,
390, 400, 410, 420, 430, 440, 450, 460, 470, 480, 490 or 500. The
intron may have a length between 80-100, 80-120, 80-140, 80-160,
80-180, 80-200, 80-250, 80-300, 80-350, 80-400, 80-450, 80-500,
200-300, 200-400, 200-500, 300-400, 300-500, or 400-500.
[0159] In one embodiment, the vector genome comprises at least one
element to enhance the transgene target specificity and expression
(See e.g., Powell et al. Viral Expression Cassette Elements to
Enhance Transgene Target Specificity and Expression in Gene
Therapy, 2015; the contents of which are herein incorporated by
reference in its entirety) such as an intron. Non-limiting examples
of introns include, MVM (67-97 bps), F.IX truncated intron 1 (300
bps), .beta.-globin SD/immunoglobulin heavy chain splice acceptor
(250 bps), adenovirus splice donor/immunoglobin splice acceptor
(500 bps), SV40 late splice donor/splice acceptor (19S/16S) (180
bps) and hybrid adenovirus splice donor/IgG splice acceptor (230
bps).
[0160] In one embodiment, the intron may be 100-500 nucleotides in
length. The intron may have a length of 80, 90, 100, 110, 120, 130,
140, 150, 160, 170, 171, 172, 173, 174, 175, 176, 177, 178, 179,
180, 190, 200, 210, 220, 230, 240, 250, 260, 270, 280, 290, 300,
310, 320, 330, 340, 350, 360, 370, 380, 390, 400, 410, 420, 430,
440, 450, 460, 470, 480, 490 or 500. The intron may have a length
between 80-100, 80-120, 80-140, 80-160, 80-180, 80-200, 80-250,
80-300, 80-350, 80-400, 80-450, 80-500, 200-300, 200-400, 200-500,
300-400, 300-500, or 400-500.
Viral Genome Component: Filler Sequence
[0161] In one embodiment, the viral genome comprises one or more
filler sequences.
[0162] In one embodiment, the viral genome comprises one or more
filler sequences in order to have the length of the viral genome be
the optimal size for packaging. As a non-limiting example, the
viral genome comprises at least one filler sequence in order to
have the length of the viral genome be about 2.3 kb. As a
non-limiting example, the viral genome comprises at least one
filler sequence in order to have the length of the viral genome be
about 4.6 kb.
[0163] In one embodiment, the viral genome is a single stranded
(ss) viral genome and comprises one or more filler sequences which
have a length about between 0.1 kb-3.8 kb, such as, but not limited
to, 0.1 kb, 0.2 kb, 0.3 kb, 0.4 kb, 0.5 kb, 0.6 kb, 0.7 kb, 0.8 kb,
0.9 kb, 1 kb, 1.1 kb, 1.2 kb, 1.3 kb, 1.4 kb, 1.5 kb, 1.6 kb, 1.7
kb, 1.8 kb, 1.9 kb, 2 kb, 2.1 kb, 2.2 kb, 2.3 kb, 2.4 kb, 2.5 kb,
2.6 kb, 2.7 kb, 2.8 kb, 2.9 kb, 3 kb, 3.1 kb, 3.2 kb, 3.3 kb, 3.4
kb, 3.5 kb, 3.6 kb, 3.7 kb, or 3.8 kb. As a non-limiting example,
the total length filler sequence in the vector genome is 3.1 kb. As
a non-limiting example, the total length filler sequence in the
vector genome is 2.7 kb. As a non-limiting example, the total
length filler sequence in the vector genome is 0.8 kb. As a
non-limiting example, the total length filler sequence in the
vector genome is 0.4 kb. As a non-limiting example, the length of
each filler sequence in the vector genome is 0.8 kb. As a
non-limiting example, the length of each filler sequence in the
vector genome is 0.4 kb.
[0164] In one embodiment, the viral genome is a self-complementary
(sc) viral genome and comprises one or more filler sequences which
have a length about between 0.1 kb-1.5 kb, such as, but not limited
to, 0.1 kb, 0.2 kb, 0.3 kb, 0.4 kb, 0.5 kb, 0.6 kb, 0.7 kb, 0.8 kb,
0.9 kb, 1 kb, 1.1 kb, 1.2 kb, 1.3 kb, 1.4 kb, or 1.5 kb. As a
non-limiting example, the total length filler sequence in the
vector genome is 0.8 kb. As a non-limiting example, the total
length filler sequence in the vector genome is 0.4 kb. As a
non-limiting example, the length of each filler sequence in the
vector genome is 0.8 kb. As a non-limiting example, the length of
each filler sequence in the vector genome is 0.4 kb
[0165] In one embodiment, the viral genome comprises any portion of
a filler sequence. The viral genome may comprise 1%, 2%, 3%, 4%,
5%, 6%, 7%, 8%, 9%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%,
55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 99% of a filler
sequence.
[0166] In one embodiment, the viral genome is a single stranded
(ss) viral genome and comprises one or more filler sequences in
order to have the length of the viral genome be about 4.6 kb. As a
non-limiting example, the viral genome comprises at least one
filler sequence and the filler sequence is located 3' to the 5' ITR
sequence. As a non-limiting example, the viral genome comprises at
least one filler sequence and the filler sequence is located 5' to
a promoter sequence. As a non-limiting example, the viral genome
comprises at least one filler sequence and the filler sequence is
located 3' to the polyadenylation signal sequence. As a
non-limiting example, the viral genome comprises at least one
filler sequence and the filler sequence is located 5' to the 3' ITR
sequence. As a non-limiting example, the viral genome comprises at
least one filler sequence, and the filler sequence is located
between two intron sequences. As a non-limiting example, the viral
genome comprises at least one filler sequence, and the filler
sequence is located within an intron sequence. As a non-limiting
example, the viral genome comprises two filler sequences, and the
first filler sequence is located 3' to the 5' ITR sequence and the
second filler sequence is located 3' to the polyadenylation signal
sequence. As a non-limiting example, the viral genome comprises two
filler sequences, and the first filler sequence is located 5' to a
promoter sequence and the second filler sequence is located 3' to
the polyadenylation signal sequence. As a non-limiting example, the
viral genome comprises two filler sequences, and the first filler
sequence is located 3' to the 5' ITR sequence and the second filler
sequence is located 5' to the 5' ITR sequence.
[0167] In one embodiment, the viral genome is a self-complementary
(sc) viral genome and comprises one or more filler sequences in
order to have the length of the viral genome be about 2.3 kb. As a
non-limiting example, the viral genome comprises at least one
filler sequence and the filler sequence is located 3' to the 5' ITR
sequence. As a non-limiting example, the viral genome comprises at
least one filler sequence and the filler sequence is located 5' to
a promoter sequence. As a non-limiting example, the viral genome
comprises at least one filler sequence and the filler sequence is
located 3' to the polyadenylation signal sequence. As a
non-limiting example, the viral genome comprises at least one
filler sequence and the filler sequence is located 5' to the 3' ITR
sequence. As a non-limiting example, the viral genome comprises at
least one filler sequence, and the filler sequence is located
between two intron sequences. As a non-limiting example, the viral
genome comprises at least one filler sequence, and the filler
sequence is located within an intron sequence. As a non-limiting
example, the viral genome comprises two filler sequences, and the
first filler sequence is located 3' to the 5' ITR sequence and the
second filler sequence is located 3' to the polyadenylation signal
sequence. As a non-limiting example, the viral genome comprises two
filler sequences, and the first filler sequence is located 5' to a
promoter sequence and the second filler sequence is located 3' to
the polyadenylation signal sequence. As a non-limiting example, the
viral genome comprises two filler sequences, and the first filler
sequence is located 3' to the 5' ITR sequence and the second filler
sequence is located 5' to the 5' ITR sequence.
[0168] In one embodiment, the viral genome may comprise one or more
filler sequences between one of more regions of the viral genome.
In one embodiment, the filler region may be located before a region
such as, but not limited to, a payload region, an inverted terminal
repeat (ITR), a promoter region, an intron region, an enhancer
region, a polyadenylation signal sequence region, a multiple
cloning site (MCS) region, and/or an exon region. In one
embodiment, the filler region may be located after a region such
as, but not limited to, a payload region, an inverted terminal
repeat (ITR), a promoter region, an intron region, an enhancer
region, a polyadenylation signal sequence region, a multiple
cloning site (MCS) region, and/or an exon region. In one
embodiment, the filler region may be located before and after a
region such as, but not limited to, a payload region, an inverted
terminal repeat (ITR), a promoter region, an intron region, an
enhancer region, a polyadenylation signal sequence region, a
multiple cloning site (MCS) region, and/or an exon region.
[0169] In one embodiment, the viral genome may comprise one or more
filler sequences which bifurcates at least one region of the viral
genome. The bifurcated region of the viral genome may comprise 1%,
2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 15%, 20%, 25%, 30%, 35%, 40%,
45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 99% of
the of the region to the 5' of the filler sequence region. As a
non-limiting example, the filler sequence may bifurcate at least
one region so that 10% of the region is located 5' to the filler
sequence and 90% of the region is located 3' to the filler
sequence. As a non-limiting example, the filler sequence may
bifurcate at least one region so that 20% of the region is located
5' to the filler sequence and 80% of the region is located 3' to
the filler sequence. As a non-limiting example, the filler sequence
may bifurcate at least one region so that 30% of the region is
located 5' to the filler sequence and 70% of the region is located
3' to the filler sequence. As a non-limiting example, the filler
sequence may bifurcate at least one region so that 40% of the
region is located 5' to the filler sequence and 60% of the region
is located 3' to the filler sequence. As a non-limiting example,
the filler sequence may bifurcate at least one region so that 50%
of the region is located 5' to the filler sequence and 50% of the
region is located 3' to the filler sequence. As a non-limiting
example, the filler sequence may bifurcate at least one region so
that 60% of the region is located 5' to the filler sequence and 40%
of the region is located 3' to the filler sequence. As a
non-limiting example, the filler sequence may bifurcate at least
one region so that 70% of the region is located 5' to the filler
sequence and 30% of the region is located 3' to the filler
sequence. As a non-limiting example, the filler sequence may
bifurcate at least one region so that 80% of the region is located
5' to the filler sequence and 20% of the region is located 3' to
the filler sequence. As a non-limiting example, the filler sequence
may bifurcate at least one region so that 90% of the region is
located 5' to the filler sequence and 10% of the region is located
3' to the filler sequence.
[0170] In one embodiment, the viral genome comprises a filler
sequence after the 5' ITR.
[0171] In one embodiment, the viral genome comprises a filler
sequence after the promoter region. In one embodiment, the viral
genome comprises a filler sequence after the payload region. In one
embodiment, the viral genome comprises a filler sequence after the
intron region. In one embodiment, the viral genome comprises a
filler sequence after the enhancer region. In one embodiment, the
viral genome comprises a filler sequence after the polyadenylation
signal sequence region. In one embodiment, the viral genome
comprises a filler sequence after the MCS region. In one
embodiment, the viral genome comprises a filler sequence after the
exon region.
[0172] In one embodiment, the viral genome comprises a filler
sequence before the promoter region. In one embodiment, the viral
genome comprises a filler sequence before the payload region. In
one embodiment, the viral genome comprises a filler sequence before
the intron region. In one embodiment, the viral genome comprises a
filler sequence before the enhancer region. In one embodiment, the
viral genome comprises a filler sequence before the polyadenylation
signal sequence region. In one embodiment, the viral genome
comprises a filler sequence before the MCS region. In one
embodiment, the viral genome comprises a filler sequence before the
exon region.
[0173] In one embodiment, the viral genome comprises a filler
sequence before the 3' ITR.
[0174] In one embodiment, a filler sequence may be located between
two regions, such as, but not limited to, the 5' ITR and the
promoter region. In one embodiment, a filler sequence may be
located between two regions, such as, but not limited to, the 5'
ITR and the payload region. In one embodiment, a filler sequence
may be located between two regions, such as, but not limited to,
the 5' ITR and the intron region. In one embodiment, a filler
sequence may be located between two regions, such as, but not
limited to, the 5' ITR and the enhancer region. In one embodiment,
a filler sequence may be located between two regions, such as, but
not limited to, the 5' ITR and the polyadenylation signal sequence
region. In one embodiment, a filler sequence may be located between
two regions, such as, but not limited to, the 5' ITR and the MCS
region.
[0175] In one embodiment, a filler sequence may be located between
two regions, such as, but not limited to, the 5' ITR and the exon
region.
[0176] In one embodiment, a filler sequence may be located between
two regions, such as, but not limited to, the promoter region and
the payload region. In one embodiment, a filler sequence may be
located between two regions, such as, but not limited to, the
promoter region and the intron region. In one embodiment, a filler
sequence may be located between two regions, such as, but not
limited to, the promoter region and the enhancer region. In one
embodiment, a filler sequence may be located between two regions,
such as, but not limited to, the promoter region and the
polyadenylation signal sequence region. In one embodiment, a filler
sequence may be located between two regions, such as, but not
limited to, the promoter region and the MCS region. In one
embodiment, a filler sequence may be located between two regions,
such as, but not limited to, the promoter region and the exon
region. In one embodiment, a filler sequence may be located between
two regions, such as, but not limited to, the promoter region and
the 3' ITR.
[0177] In one embodiment, a filler sequence may be located between
two regions, such as, but not limited to, the payload region and
the intron region. In one embodiment, a filler sequence may be
located between two regions, such as, but not limited to, the
payload region and the enhancer region. In one embodiment, a filler
sequence may be located between two regions, such as, but not
limited to, the payload region and the polyadenylation signal
sequence region. In one embodiment, a filler sequence may be
located between two regions, such as, but not limited to, the
payload region and the MCS region. In one embodiment, a filler
sequence may be located between two regions, such as, but not
limited to, the payload region and the exon region.
[0178] In one embodiment, a filler sequence may be located between
two regions, such as, but not limited to, the payload region and
the 3' ITR.
[0179] In one embodiment, a filler sequence may be located between
two regions, such as, but not limited to, the intron region and the
enhancer region. In one embodiment, a filler sequence may be
located between two regions, such as, but not limited to, the
intron region and the polyadenylation signal sequence region. In
one embodiment, a filler sequence may be located between two
regions, such as, but not limited to, the intron region and the MCS
region. In one embodiment, a filler sequence may be located between
two regions, such as, but not limited to, the intron region and the
exon region. In one embodiment, a filler sequence may be located
between two regions, such as, but not limited to, the intron region
and the 3' ITR. In one embodiment, a filler sequence may be located
between two regions, such as, but not limited to, the enhancer
region and the polyadenylation signal sequence region. In one
embodiment, a filler sequence may be located between two regions,
such as, but not limited to, the enhancer region and the MCS
region. In one embodiment, a filler sequence may be located between
two regions, such as, but not limited to, the enhancer region and
the exon region. In one embodiment, a filler sequence may be
located between two regions, such as, but not limited to, the
enhancer region and the 3' ITR.
[0180] In one embodiment, a filler sequence may be located between
two regions, such as, but not limited to, the polyadenylation
signal sequence region and the MCS region. In one embodiment, a
filler sequence may be located between two regions, such as, but
not limited to, the polyadenylation signal sequence region and the
exon region. In one embodiment, a filler sequence may be located
between two regions, such as, but not limited to, the
polyadenylation signal sequence region and the 3' ITR.
[0181] In one embodiment, a filler sequence may be located between
two regions, such as, but not limited to, the MCS region and the
exon region. In one embodiment, a filler sequence may be located
between two regions, such as, but not limited to, the MCS region
and the 3' ITR.
[0182] In one embodiment, a filler sequence may be located between
two regions, such as, but not limited to, the exon region and the
3' ITR.
[0183] In one embodiment, a viral genome may comprise two filler
sequences, the first filler sequence may be located between the 5'
ITR and promoter region, and the second filler sequence may be
located between the promoter region and payload region. In one
embodiment, a viral genome may comprise two filler sequences, the
first filler sequence may be located between the 5' ITR and
promoter region, and the second filler sequence may be located
between the promoter region and intron region. In one embodiment, a
viral genome may comprise two filler sequences, the first filler
sequence may be located between the 5' ITR and promoter region, and
the second filler sequence may be located between the promoter
region and enhancer region. In one embodiment, a viral genome may
comprise two filler sequences, the first filler sequence may be
located between the 5' ITR and promoter region, and the second
filler sequence may be located between the promoter region and
polyadenylation signal sequence region. In one embodiment, a viral
genome may comprise two filler sequences, the first filler sequence
may be located between the 5' ITR and promoter region, and the
second filler sequence may be located between the promoter region
and MCS region. In one embodiment, a viral genome may comprise two
filler sequences, the first filler sequence may be located between
the 5' ITR and promoter region, and the second filler sequence may
be located between the promoter region and exon region. In one
embodiment, a viral genome may comprise two filler sequences, the
first filler sequence may be located between the 5' ITR and
promoter region, and the second filler sequence may be located
between the promoter region and 3' ITR. In one embodiment, a viral
genome may comprise two filler sequences, the first filler sequence
may be located between the 5' ITR and promoter region, and the
second filler sequence may be located between the payload region
and intron region. In one embodiment, a viral genome may comprise
two filler sequences, the first filler sequence may be located
between the 5' ITR and promoter region, and the second filler
sequence may be located between the payload region and enhancer
region. In one embodiment, a viral genome may comprise two filler
sequences, the first filler sequence may be located between the 5'
ITR and promoter region, and the second filler sequence may be
located between the payload region and polyadenylation signal
sequence region. In one embodiment, a viral genome may comprise two
filler sequences, the first filler sequence may be located between
the 5' ITR and promoter region, and the second filler sequence may
be located between the payload region and MCS region. In one
embodiment, a viral genome may comprise two filler sequences, the
first filler sequence may be located between the 5' ITR and
promoter region, and the second filler sequence may be located
between the payload region and exon region. In one embodiment, a
viral genome may comprise two filler sequences, the first filler
sequence may be located between the 5' ITR and promoter region, and
the second filler sequence may be located between the payload
region and 3' ITR. In one embodiment, a viral genome may comprise
two filler sequences, the first filler sequence may be located
between the 5' ITR and promoter region, and the second filler
sequence may be located between the intron region and enhancer
region. In one embodiment, a viral genome may comprise two filler
sequences, the first filler sequence may be located between the 5'
ITR and promoter region, and the second filler sequence may be
located between the intron region and polyadenylation signal
sequence region. In one embodiment, a viral genome may comprise two
filler sequences, the first filler sequence may be located between
the 5' ITR and promoter region, and the second filler sequence may
be located between the intron region and MCS region. In one
embodiment, a viral genome may comprise two filler sequences, the
first filler sequence may be located between the 5' ITR and
promoter region, and the second filler sequence may be located
between the intron region and exon region. In one embodiment, a
viral genome may comprise two filler sequences, the first filler
sequence may be located between the 5' ITR and promoter region, and
the second filler sequence may be located between the intron region
and 3' ITR. In one embodiment, a viral genome may comprise two
filler sequences, the first filler sequence may be located between
the 5' ITR and promoter region, and the second filler sequence may
be located between the enhancer region and polyadenylation signal
sequence region. In one embodiment, a viral genome may comprise two
filler sequences, the first filler sequence may be located between
the 5' ITR and promoter region, and the second filler sequence may
be located between the enhancer region and MCS region. In one
embodiment, a viral genome may comprise two filler sequences, the
first filler sequence may be located between the 5' ITR and
promoter region, and the second filler sequence may be located
between the enhancer region and exon region. In one embodiment, a
viral genome may comprise two filler sequences, the first filler
sequence may be located between the 5' ITR and promoter region, and
the second filler sequence may be located between the enhancer
region and 3' ITR. In one embodiment, a viral genome may comprise
two filler sequences, the first filler sequence may be located
between the 5' ITR and promoter region, and the second filler
sequence may be located between the polyadenylation signal sequence
region and MCS region. In one embodiment, a viral genome may
comprise two filler sequences, the first filler sequence may be
located between the 5' ITR and promoter region, and the second
filler sequence may be located between the polyadenylation signal
sequence region and exon region. In one embodiment, a viral genome
may comprise two filler sequences, the first filler sequence may be
located between the 5' ITR and promoter region, and the second
filler sequence may be located between the polyadenylation signal
sequence region and 3' ITR. In one embodiment, a viral genome may
comprise two filler sequences, the first filler sequence may be
located between the 5' ITR and promoter region, and the second
filler sequence may be located between the MCS region and exon
region. In one embodiment, a viral genome may comprise two filler
sequences, the first filler sequence may be located between the 5'
ITR and promoter region, and the second filler sequence may be
located between the MCS region and 3' ITR. In one embodiment, a
viral genome may comprise two filler sequences, the first filler
sequence may be located between the 5' ITR and promoter region, and
the second filler sequence may be located between the exon region
and 3' ITR.
[0184] In one embodiment, a viral genome may comprise two filler
sequences, the first filler sequence may be located between the 5'
ITR and payload region, and the second filler sequence may be
located between the promoter region and payload region. In one
embodiment, a viral genome may comprise two filler sequences, the
first filler sequence may be located between the 5' ITR and payload
region, and the second filler sequence may be located between the
promoter region and intron region. In one embodiment, a viral
genome may comprise two filler sequences, the first filler sequence
may be located between the 5' ITR and payload region, and the
second filler sequence may be located between the promoter region
and enhancer region. In one embodiment, a viral genome may comprise
two filler sequences, the first filler sequence may be located
between the 5' ITR and payload region, and the second filler
sequence may be located between the promoter region and
polyadenylation signal sequence region. In one embodiment, a viral
genome may comprise two filler sequences, the first filler sequence
may be located between the 5' ITR and payload region, and the
second filler sequence may be located between the promoter region
and MCS region. In one embodiment, a viral genome may comprise two
filler sequences, the first filler sequence may be located between
the 5' ITR and payload region, and the second filler sequence may
be located between the promoter region and exon region. In one
embodiment, a viral genome may comprise two filler sequences, the
first filler sequence may be located between the 5' ITR and payload
region, and the second filler sequence may be located between the
promoter region and 3' ITR. In one embodiment, a viral genome may
comprise two filler sequences, the first filler sequence may be
located between the 5' ITR and payload region, and the second
filler sequence may be located between the payload region and
intron region. In one embodiment, a viral genome may comprise two
filler sequences, the first filler sequence may be located between
the 5' ITR and payload region, and the second filler sequence may
be located between the payload region and enhancer region. In one
embodiment, a viral genome may comprise two filler sequences, the
first filler sequence may be located between the 5' ITR and payload
region, and the second filler sequence may be located between the
payload region and polyadenylation signal sequence region. In one
embodiment, a viral genome may comprise two filler sequences, the
first filler sequence may be located between the 5' ITR and payload
region, and the second filler sequence may be located between the
payload region and MCS region. In one embodiment, a viral genome
may comprise two filler sequences, the first filler sequence may be
located between the 5' ITR and payload region, and the second
filler sequence may be located between the payload region and exon
region. In one embodiment, a viral genome may comprise two filler
sequences, the first filler sequence may be located between the 5'
ITR and payload region, and the second filler sequence may be
located between the payload region and 3' ITR. In one embodiment, a
viral genome may comprise two filler sequences, the first filler
sequence may be located between the 5' ITR and payload region, and
the second filler sequence may be located between the intron region
and enhancer region. In one embodiment, a viral genome may comprise
two filler sequences, the first filler sequence may be located
between the 5' ITR and payload region, and the second filler
sequence may be located between the intron region and
polyadenylation signal sequence region. In one embodiment, a viral
genome may comprise two filler sequences, the first filler sequence
may be located between the 5' ITR and payload region, and the
second filler sequence may be located between the intron region and
MCS region. In one embodiment, a viral genome may comprise two
filler sequences, the first filler sequence may be located between
the 5' ITR and payload region, and the second filler sequence may
be located between the intron region and exon region. In one
embodiment, a viral genome may comprise two filler sequences, the
first filler sequence may be located between the 5' ITR and payload
region, and the second filler sequence may be located between the
intron region and 3' ITR. In one embodiment, a viral genome may
comprise two filler sequences, the first filler sequence may be
located between the 5' ITR and payload region, and the second
filler sequence may be located between the enhancer region and
polyadenylation signal sequence region. In one embodiment, a viral
genome may comprise two filler sequences, the first filler sequence
may be located between the 5' ITR and payload region, and the
second filler sequence may be located between the enhancer region
and MCS region. In one embodiment, a viral genome may comprise two
filler sequences, the first filler sequence may be located between
the 5' ITR and payload region, and the second filler sequence may
be located between the enhancer region and exon region. In one
embodiment, a viral genome may comprise two filler sequences, the
first filler sequence may be located between the 5' ITR and payload
region, and the second filler sequence may be located between the
enhancer region and 3' ITR. In one embodiment, a viral genome may
comprise two filler sequences, the first filler sequence may be
located between the 5' ITR and payload region, and the second
filler sequence may be located between the polyadenylation signal
sequence region and MCS region. In one embodiment, a viral genome
may comprise two filler sequences, the first filler sequence may be
located between the 5' ITR and payload region, and the second
filler sequence may be located between the polyadenylation signal
sequence region and exon region. In one embodiment, a viral genome
may comprise two filler sequences, the first filler sequence may be
located between the 5' ITR and payload region, and the second
filler sequence may be located between the polyadenylation signal
sequence region and 3' ITR. In one embodiment, a viral genome may
comprise two filler sequences, the first filler sequence may be
located between the 5' ITR and payload region, and the second
filler sequence may be located between the MCS region and exon
region. In one embodiment, a viral genome may comprise two filler
sequences, the first filler sequence may be located between the 5'
ITR and payload region, and the second filler sequence may be
located between the MCS region and 3' ITR. In one embodiment, a
viral genome may comprise two filler sequences, the first filler
sequence may be located between the 5' ITR and payload region, and
the second filler sequence may be located between the exon region
and 3' ITR.
[0185] In one embodiment, a viral genome may comprise two filler
sequences, the first filler sequence may be located between the 5'
ITR and intron region, and the second filler sequence may be
located between the promoter region and payload region. In one
embodiment, a viral genome may comprise two filler sequences, the
first filler sequence may be located between the 5' ITR and intron
region, and the second filler sequence may be located between the
promoter region and intron region. In one embodiment, a viral
genome may comprise two filler sequences, the first filler sequence
may be located between the 5' ITR and intron region, and the second
filler sequence may be located between the promoter region and
enhancer region. In one embodiment, a viral genome may comprise two
filler sequences, the first filler sequence may be located between
the 5' ITR and intron region, and the second filler sequence may be
located between the promoter region and polyadenylation signal
sequence region. In one embodiment, a viral genome may comprise two
filler sequences, the first filler sequence may be located between
the 5' ITR and intron region, and the second filler sequence may be
located between the promoter region and MCS region. In one
embodiment, a viral genome may comprise two filler sequences, the
first filler sequence may be located between the 5' ITR and intron
region, and the second filler sequence may be located between the
promoter region and exon region. In one embodiment, a viral genome
may comprise two filler sequences, the first filler sequence may be
located between the 5' ITR and intron region, and the second filler
sequence may be located between the promoter region and 3' ITR. In
one embodiment, a viral genome may comprise two filler sequences,
the first filler sequence may be located between the 5' ITR and
intron region, and the second filler sequence may be located
between the payload region and intron region. In one embodiment, a
viral genome may comprise two filler sequences, the first filler
sequence may be located between the 5' ITR and intron region, and
the second filler sequence may be located between the payload
region and enhancer region. In one embodiment, a viral genome may
comprise two filler sequences, the first filler sequence may be
located between the 5' ITR and intron region, and the second filler
sequence may be located between the payload region and
polyadenylation signal sequence region. In one embodiment, a viral
genome may comprise two filler sequences, the first filler sequence
may be located between the 5' ITR and intron region, and the second
filler sequence may be located between the payload region and MCS
region. In one embodiment, a viral genome may comprise two filler
sequences, the first filler sequence may be located between the 5'
ITR and intron region, and the second filler sequence may be
located between the payload region and exon region. In one
embodiment, a viral genome may comprise two filler sequences, the
first filler sequence may be located between the 5' ITR and intron
region, and the second filler sequence may be located between the
payload region and 3' ITR. In one embodiment, a viral genome may
comprise two filler sequences, the first filler sequence may be
located between the 5' ITR and intron region, and the second filler
sequence may be located between the intron region and enhancer
region. In one embodiment, a viral genome may comprise two filler
sequences, the first filler sequence may be located between the 5'
ITR and intron region, and the second filler sequence may be
located between the intron region and polyadenylation signal
sequence region. In one embodiment, a viral genome may comprise two
filler sequences, the first filler sequence may be located between
the 5' ITR and intron region, and the second filler sequence may be
located between the intron region and MCS region. In one
embodiment, a viral genome may comprise two filler sequences, the
first filler sequence may be located between the 5' ITR and intron
region, and the second filler sequence may be located between the
intron region and exon region. In one embodiment, a viral genome
may comprise two filler sequences, the first filler sequence may be
located between the 5' ITR and intron region, and the second filler
sequence may be located between the intron region and 3' ITR. In
one embodiment, a viral genome may comprise two filler sequences,
the first filler sequence may be located between the 5' ITR and
intron region, and the second filler sequence may be located
between the enhancer region and polyadenylation signal sequence
region. In one embodiment, a viral genome may comprise two filler
sequences, the first filler sequence may be located between the 5'
ITR and intron region, and the second filler sequence may be
located between the enhancer region and MCS region. In one
embodiment, a viral genome may comprise two filler sequences, the
first filler sequence may be located between the 5' ITR and intron
region, and the second filler sequence may be located between the
enhancer region and exon region. In one embodiment, a viral genome
may comprise two filler sequences, the first filler sequence may be
located between the 5' ITR and intron region, and the second filler
sequence may be located between the enhancer region and 3' ITR. In
one embodiment, a viral genome may comprise two filler sequences,
the first filler sequence may be located between the 5' ITR and
intron region, and the second filler sequence may be located
between the polyadenylation signal sequence region and MCS region.
In one embodiment, a viral genome may comprise two filler
sequences, the first filler sequence may be located between the 5'
ITR and intron region, and the second filler sequence may be
located between the polyadenylation signal sequence region and exon
region. In one embodiment, a viral genome may comprise two filler
sequences, the first filler sequence may be located between the 5'
ITR and intron region, and the second filler sequence may be
located between the polyadenylation signal sequence region and 3'
ITR. In one embodiment, a viral genome may comprise two filler
sequences, the first filler sequence may be located between the 5'
ITR and intron region, and the second filler sequence may be
located between the MCS region and exon region. In one embodiment,
a viral genome may comprise two filler sequences, the first filler
sequence may be located between the 5' ITR and intron region, and
the second filler sequence may be located between the MCS region
and 3' ITR. In one embodiment, a viral genome may comprise two
filler sequences, the first filler sequence may be located between
the 5' ITR and intron region, and the second filler sequence may be
located between the exon region and 3' ITR.
[0186] In one embodiment, a viral genome may comprise two filler
sequences, the first filler sequence may be located between the 5'
ITR and enhancer region, and the second filler sequence may be
located between the promoter region and payload region. In one
embodiment, a viral genome may comprise two filler sequences, the
first filler sequence may be located between the 5' ITR and
enhancer region, and the second filler sequence may be located
between the promoter region and intron region. In one embodiment, a
viral genome may comprise two filler sequences, the first filler
sequence may be located between the 5' ITR and enhancer region, and
the second filler sequence may be located between the promoter
region and enhancer region. In one embodiment, a viral genome may
comprise two filler sequences, the first filler sequence may be
located between the 5' ITR and enhancer region, and the second
filler sequence may be located between the promoter region and
polyadenylation signal sequence region. In one embodiment, a viral
genome may comprise two filler sequences, the first filler sequence
may be located between the 5' ITR and enhancer region, and the
second filler sequence may be located between the promoter region
and MCS region. In one embodiment, a viral genome may comprise two
filler sequences, the first filler sequence may be located between
the 5' ITR and enhancer region, and the second filler sequence may
be located between the promoter region and exon region. In one
embodiment, a viral genome may comprise two filler sequences, the
first filler sequence may be located between the 5' ITR and
enhancer region, and the second filler sequence may be located
between the promoter region and 3' ITR. In one embodiment, a viral
genome may comprise two filler sequences, the first filler sequence
may be located between the 5' ITR and enhancer region, and the
second filler sequence may be located between the payload region
and intron region. In one embodiment, a viral genome may comprise
two filler sequences, the first filler sequence may be located
between the 5' ITR and enhancer region, and the second filler
sequence may be located between the payload region and enhancer
region. In one embodiment, a viral genome may comprise two filler
sequences, the first filler sequence may be located between the 5'
ITR and enhancer region, and the second filler sequence may be
located between the payload region and polyadenylation signal
sequence region. In one embodiment, a viral genome may comprise two
filler sequences, the first filler sequence may be located between
the 5' ITR and enhancer region, and the second filler sequence may
be located between the payload region and MCS region. In one
embodiment, a viral genome may comprise two filler sequences, the
first filler sequence may be located between the 5' ITR and
enhancer region, and the second filler sequence may be located
between the payload region and exon region. In one embodiment, a
viral genome may comprise two filler sequences, the first filler
sequence may be located between the 5' ITR and enhancer region, and
the second filler sequence may be located between the payload
region and 3' ITR. In one embodiment, a viral genome may comprise
two filler sequences, the first filler sequence may be located
between the 5' ITR and enhancer region, and the second filler
sequence may be located between the intron region and enhancer
region. In one embodiment, a viral genome may comprise two filler
sequences, the first filler sequence may be located between the 5'
ITR and enhancer region, and the second filler sequence may be
located between the intron region and polyadenylation signal
sequence region. In one embodiment, a viral genome may comprise two
filler sequences, the first filler sequence may be located between
the 5' ITR and enhancer region, and the second filler sequence may
be located between the intron region and MCS region. In one
embodiment, a viral genome may comprise two filler sequences, the
first filler sequence may be located between the 5' ITR and
enhancer region, and the second filler sequence may be located
between the intron region and exon region. In one embodiment, a
viral genome may comprise two filler sequences, the first filler
sequence may be located between the 5' ITR and enhancer region, and
the second filler sequence may be located between the intron region
and 3' ITR. In one embodiment, a viral genome may comprise two
filler sequences, the first filler sequence may be located between
the 5' ITR and enhancer region, and the second filler sequence may
be located between the enhancer region and polyadenylation signal
sequence region. In one embodiment, a viral genome may comprise two
filler sequences, the first filler sequence may be located between
the 5' ITR and enhancer region, and the second filler sequence may
be located between the enhancer region and MCS region. In one
embodiment, a viral genome may comprise two filler sequences, the
first filler sequence may be located between the 5' ITR and
enhancer region, and the second filler sequence may be located
between the enhancer region and exon region. In one embodiment, a
viral genome may comprise two filler sequences, the first filler
sequence may be located between the 5' ITR and enhancer region, and
the second filler sequence may be located between the enhancer
region and 3' ITR. In one embodiment, a viral genome may comprise
two filler sequences, the first filler sequence may be located
between the 5' ITR and enhancer region, and the second filler
sequence may be located between the polyadenylation signal sequence
region and MCS region. In one embodiment, a viral genome may
comprise two filler sequences, the first filler sequence may be
located between the 5' ITR and enhancer region, and the second
filler sequence may be located between the polyadenylation signal
sequence region and exon region. In one embodiment, a viral genome
may comprise two filler sequences, the first filler sequence may be
located between the 5' ITR and enhancer region, and the second
filler sequence may be located between the polyadenylation signal
sequence region and 3' ITR. In one embodiment, a viral genome may
comprise two filler sequences, the first filler sequence may be
located between the 5' ITR and enhancer region, and the second
filler sequence may be located between the MCS region and exon
region. In one embodiment, a viral genome may comprise two filler
sequences, the first filler sequence may be located between the 5'
ITR and enhancer region, and the second filler sequence may be
located between the MCS region and 3' ITR. In one embodiment, a
viral genome may comprise two filler sequences, the first filler
sequence may be located between the 5' ITR and enhancer region, and
the second filler sequence may be located between the exon region
and 3' ITR.
[0187] In one embodiment, a viral genome may comprise two filler
sequences, the first filler sequence may be located between the 5'
ITR and polyadenylation signal sequence region, and the second
filler sequence may be located between the promoter region and
payload region. In one embodiment, a viral genome may comprise two
filler sequences, the first filler sequence may be located between
the 5' ITR and polyadenylation signal sequence region, and the
second filler sequence may be located between the promoter region
and intron region. In one embodiment, a viral genome may comprise
two filler sequences, the first filler sequence may be located
between the 5' ITR and polyadenylation signal sequence region, and
the second filler sequence may be located between the promoter
region and enhancer region. In one embodiment, a viral genome may
comprise two filler sequences, the first filler sequence may be
located between the 5' ITR and polyadenylation signal sequence
region, and the second filler sequence may be located between the
promoter region and polyadenylation signal sequence region. In one
embodiment, a viral genome may comprise two filler sequences, the
first filler sequence may be located between the 5' ITR and
promoter region and MCS region. In one embodiment, a viral genome
may comprise two filler sequences, the first filler sequence may be
located between the 5' ITR and polyadenylation signal sequence
region, and the second filler sequence may be located between the
promoter region and exon region. In one embodiment, a viral genome
may comprise two filler sequences, the first filler sequence may be
located between the 5' ITR and polyadenylation signal sequence
region, and the second filler sequence may be located between the
promoter region and 3' ITR. In one embodiment, a viral genome may
comprise two filler sequences, the first filler sequence may be
located between the 5' ITR and polyadenylation signal sequence
region, and the second filler sequence may be located between the
payload region and intron region. In one embodiment, a viral genome
may comprise two filler sequences, the first filler sequence may be
located between the 5' ITR and polyadenylation signal sequence
region, and the second filler sequence may be located between the
payload region and enhancer region. In one embodiment, a viral
genome may comprise two filler sequences, the first filler sequence
may be located between the 5' ITR and polyadenylation signal
sequence region, and the second filler sequence may be located
between the payload region and polyadenylation signal sequence
region. In one embodiment, a viral genome may comprise two filler
sequences, the first filler sequence may be located between the 5'
ITR and polyadenylation signal sequence region, and the second
filler sequence may be located between the payload region and MCS
region. In one embodiment, a viral genome may comprise two filler
sequences, the first filler sequence may be located between the 5'
ITR and polyadenylation signal sequence region, and the second
filler sequence may be located between the payload region and exon
region. In one embodiment, a viral genome may comprise two filler
sequences, the first filler sequence may be located between the 5'
ITR and polyadenylation signal sequence region, and the second
filler sequence may be located between the payload region and 3'
ITR. In one embodiment, a viral genome may comprise two filler
sequences, the first filler sequence may be located between the 5'
ITR and polyadenylation signal sequence region, and the second
filler sequence may be located between the intron region and
enhancer region. In one embodiment, a viral genome may comprise two
filler sequences, the first filler sequence may be located between
the 5' ITR and polyadenylation signal sequence region, and the
second filler sequence may be located between the intron region and
polyadenylation signal sequence region. In one embodiment, a viral
genome may comprise two filler sequences, the first filler sequence
may be located between the 5' ITR and intron region and MCS region.
In one embodiment, a viral genome may comprise two filler
sequences, the first filler sequence may be located between the 5'
ITR and polyadenylation signal sequence region, and the second
filler sequence may be located between the intron region and exon
region. In one embodiment, a viral genome may comprise two filler
sequences, the first filler sequence may be located between the 5'
ITR and polyadenylation signal sequence region, and the second
filler sequence may be located between the intron region and 3'
ITR. In one embodiment, a viral genome may comprise two filler
sequences, the first filler sequence may be located between the 5'
ITR and polyadenylation signal sequence region, and the second
filler sequence may be located between the enhancer region and
polyadenylation signal sequence region. In one embodiment, a viral
genome may comprise two filler sequences, the first filler sequence
may be located between the 5' ITR and polyadenylation signal
sequence region, and the second filler sequence may be located
between the enhancer region and MCS region. In one embodiment, a
viral genome may comprise two filler sequences, the first filler
sequence may be located between the 5' ITR and polyadenylation
signal sequence region, and the second filler sequence may be
located between the enhancer region and exon region. In one
embodiment, a viral genome may comprise two filler sequences, the
first filler sequence may be located between the 5' ITR and
polyadenylation signal sequence region, and the second filler
sequence may be located between the enhancer region and 3' ITR. In
one embodiment, a viral genome may comprise two filler sequences,
the first filler sequence may be located between the 5' ITR and
polyadenylation signal sequence region, and the second filler
sequence may be located between the polyadenylation signal sequence
region and MCS region. In one embodiment, a viral genome may
comprise two filler sequences, the first filler sequence may be
located between the 5' ITR and polyadenylation signal sequence
region, and the second filler sequence may be located between the
polyadenylation signal sequence region and exon region. In one
embodiment, a viral genome may comprise two filler sequences, the
first filler sequence may be located between the 5' ITR and
polyadenylation signal sequence region, and the second filler
sequence may be located between the polyadenylation signal sequence
region and 3' ITR. In one embodiment, a viral genome may comprise
two filler sequences, the first filler sequence may be located
between the 5' ITR and polyadenylation signal sequence region, and
the second filler sequence may be located between the MCS region
and exon region. In one embodiment, a viral genome may comprise two
filler sequences, the first filler sequence may be located between
the 5' ITR and polyadenylation signal sequence region, and the
second filler sequence may be located between the MCS region and 3'
ITR. In one embodiment, a viral genome may comprise two filler
sequences, the first filler sequence may be located between the 5'
ITR and polyadenylation signal sequence region, and the second
filler sequence may be located between the exon region and 3'
ITR.
[0188] In one embodiment, a viral genome may comprise two filler
sequences, the first filler sequence may be located between the 5'
ITR and MCS region, and the second filler sequence may be located
between the promoter region and payload region. In one embodiment,
a viral genome may comprise two filler sequences, the first filler
sequence may be located between the 5' ITR and MCS region, and the
second filler sequence may be located between the promoter region
and intron region. In one embodiment, a viral genome may comprise
two filler sequences, the first filler sequence may be located
between the 5' ITR and MCS region, and the second filler sequence
may be located between the promoter region and enhancer region. In
one embodiment, a viral genome may comprise two filler sequences,
the first filler sequence may be located between the 5' ITR and MCS
region, and the second filler sequence may be located between the
promoter region and polyadenylation signal sequence region. In one
embodiment, a viral genome may comprise two filler sequences, the
first filler sequence may be located between the 5' ITR and MCS
region, and the second filler sequence may be located between the
promoter region and MCS region. In one embodiment, a viral genome
may comprise two filler sequences, the first filler sequence may be
located between the 5' ITR and MCS region, and the second filler
sequence may be located between the promoter region and exon
region. In one embodiment, a viral genome may comprise two filler
sequences, the first filler sequence may be located between the 5'
ITR and MCS region, and the second filler sequence may be located
between the promoter region and 3' ITR. In one embodiment, a viral
genome may comprise two filler sequences, the first filler sequence
may be located between the 5' ITR and MCS region, and the second
filler sequence may be located between the payload region and
intron region. In one embodiment, a viral genome may comprise two
filler sequences, the first filler sequence may be located between
the 5' ITR and MCS region, and the second filler sequence may be
located between the payload region and enhancer region. In one
embodiment, a viral genome may comprise two filler sequences, the
first filler sequence may be located between the 5' ITR and MCS
region, and the second filler sequence may be located between the
payload region and polyadenylation signal sequence region. In one
embodiment, a viral genome may comprise two filler sequences, the
first filler sequence may be located between the 5' ITR and MCS
region, and the second filler sequence may be located between the
payload region and MCS region. In one embodiment, a viral genome
may comprise two filler sequences, the first filler sequence may be
located between the 5' ITR and MCS region, and the second filler
sequence may be located between the payload region and exon region.
In one embodiment, a viral genome may comprise two filler
sequences, the first filler sequence may be located between the 5'
ITR and MCS region, and the second filler sequence may be located
between the payload region and 3' ITR. In one embodiment, a viral
genome may comprise two filler sequences, the first filler sequence
may be located between the 5' ITR and MCS region, and the second
filler sequence may be located between the intron region and
enhancer region. In one embodiment, a viral genome may comprise two
filler sequences, the first filler sequence may be located between
the 5' ITR and MCS region, and the second filler sequence may be
located between the intron region and polyadenylation signal
sequence region. In one embodiment, a viral genome may comprise two
filler sequences, the first filler sequence may be located between
the 5' ITR and MCS region, and the second filler sequence may be
located between the intron region and MCS region. In one
embodiment, a viral genome may comprise two filler sequences, the
first filler sequence may be located between the 5' ITR and MCS
region, and the second filler sequence may be located between the
intron region and exon region. In one embodiment, a viral genome
may comprise two filler sequences, the first filler sequence may be
located between the 5' ITR and MCS region, and the second filler
sequence may be located between the intron region and 3' ITR. In
one embodiment, a viral genome may comprise two filler sequences,
the first filler sequence may be located between the 5' ITR and MCS
region, and the second filler sequence may be located between the
enhancer region and polyadenylation signal sequence region. In one
embodiment, a viral genome may comprise two filler sequences, the
first filler sequence may be located between the 5' ITR and MCS
region, and the second filler sequence may be located between the
enhancer region and MCS region. In one embodiment, a viral genome
may comprise two filler sequences, the first filler sequence may be
located between the 5' ITR and MCS region, and the second filler
sequence may be located between the enhancer region and exon
region. In one embodiment, a viral genome may comprise two filler
sequences, the first filler sequence may be located between the 5'
ITR and MCS region, and the second filler sequence may be located
between the enhancer region and 3' ITR. In one embodiment, a viral
genome may comprise two filler sequences, the first filler sequence
may be located between the 5' ITR and MCS region, and the second
filler sequence may be located between the polyadenylation signal
sequence region and MCS region. In one embodiment, a viral genome
may comprise two filler sequences, the first filler sequence may be
located between the 5' ITR and MCS region, and the second filler
sequence may be located between the polyadenylation signal sequence
region and exon region. In one embodiment, a viral genome may
comprise two filler sequences, the first filler sequence may be
located between the 5' ITR and MCS region, and the second filler
sequence may be located between the polyadenylation signal sequence
region and 3' ITR. In one embodiment, a viral genome may comprise
two filler sequences, the first filler sequence may be located
between the 5' ITR and MCS region, and the second filler sequence
may be located between the MCS region and exon region. In one
embodiment, a viral genome may comprise two filler sequences, the
first filler sequence may be located between the 5' ITR and MCS
region, and the second filler sequence may be located between the
MCS region and 3' ITR. In one embodiment, a viral genome may
comprise two filler sequences, the first filler sequence may be
located between the 5' ITR and MCS region, and the second filler
sequence may be located between the exon region and 3' ITR.
[0189] In one embodiment, a viral genome may comprise two filler
sequences, the first filler sequence may be located between the 5'
ITR and exon region, and the second filler sequence may be located
between the promoter region and payload region. In one embodiment,
a viral genome may comprise two filler sequences, the first filler
sequence may be located between the 5' ITR and exon region, and the
second filler sequence may be located between the promoter region
and intron region. In one embodiment, a viral genome may comprise
two filler sequences, the first filler sequence may be located
between the 5' ITR and exon region, and the second filler sequence
may be located between the promoter region and enhancer region. In
one embodiment, a viral genome may comprise two filler sequences,
the first filler sequence may be located between the 5' ITR and
exon region, and the second filler sequence may be located between
the promoter region and polyadenylation signal sequence region. In
one embodiment, a viral genome may comprise two filler sequences,
the first filler sequence may be located between the 5' ITR and
exon region, and the second filler sequence may be located between
the promoter region and MCS region. In one embodiment, a viral
genome may comprise two filler sequences, the first filler sequence
may be located between the 5' ITR and exon region, and the second
filler sequence may be located between the promoter region and exon
region. In one embodiment, a viral genome may comprise two filler
sequences, the first filler sequence may be located between the 5'
ITR and exon region, and the second filler sequence may be located
between the promoter region and 3' ITR. In one embodiment, a viral
genome may comprise two filler sequences, the first filler sequence
may be located between the 5' ITR and exon region, and the second
filler sequence may be located between the payload region and
intron region. In one embodiment, a viral genome may comprise two
filler sequences, the first filler sequence may be located between
the 5' ITR and exon region, and the second filler sequence may be
located between the payload region and enhancer region. In one
embodiment, a viral genome may comprise two filler sequences, the
first filler sequence may be located between the 5' ITR and exon
region, and the second filler sequence may be located between the
payload region and polyadenylation signal sequence region. In one
embodiment, a viral genome may comprise two filler sequences, the
first filler sequence may be located between the 5' ITR and exon
region, and the second filler sequence may be located between the
payload region and MCS region. In one embodiment, a viral genome
may comprise two filler sequences, the first filler sequence may be
located between the 5' ITR and exon region, and the second filler
sequence may be located between the payload region and exon region.
In one embodiment, a viral genome may comprise two filler
sequences, the first filler sequence may be located between the 5'
ITR and exon region, and the second filler sequence may be located
between the payload region and 3' ITR. In one embodiment, a viral
genome may comprise two filler sequences, the first filler sequence
may be located between the 5' ITR and exon region, and the second
filler sequence may be located between the intron region and
enhancer region. In one embodiment, a viral genome may comprise two
filler sequences, the first filler sequence may be located between
the 5' ITR and exon region, and the second filler sequence may be
located between the intron region and polyadenylation signal
sequence region. In one embodiment, a viral genome may comprise two
filler sequences, the first filler sequence may be located between
the 5' ITR and exon region, and the second filler sequence may be
located between the intron region and MCS region. In one
embodiment, a viral genome may comprise two filler sequences, the
first filler sequence may be located between the 5' ITR and exon
region, and the second filler sequence may be located between the
intron region and exon region. In one embodiment, a viral genome
may comprise two filler sequences, the first filler sequence may be
located between the 5' ITR and exon region, and the second filler
sequence may be located between the intron region and 3' ITR. In
one embodiment, a viral genome may comprise two filler sequences,
the first filler sequence may be located between the 5' ITR and
exon region, and the second filler sequence may be located between
the enhancer region and polyadenylation signal sequence region. In
one embodiment, a viral genome may comprise two filler sequences,
the first filler sequence may be located between the 5' ITR and
exon region, and the second filler sequence may be located between
the enhancer region and MCS region. In one embodiment, a viral
genome may comprise two filler sequences, the first filler sequence
may be located between the 5' ITR and exon region, and the second
filler sequence may be located between the enhancer region and exon
region. In one embodiment, a viral genome may comprise two filler
sequences, the first filler sequence may be located between the 5'
ITR and exon region, and the second filler sequence may be located
between the enhancer region and 3' ITR. In one embodiment, a viral
genome may comprise two filler sequences, the first filler sequence
may be located between the 5' ITR and exon region, and the second
filler sequence may be located between the polyadenylation signal
sequence region and MCS region. In one embodiment, a viral genome
may comprise two filler sequences, the first filler sequence may be
located between the 5' ITR and exon region, and the second filler
sequence may be located between the polyadenylation signal sequence
region and exon region. In one embodiment, a viral genome may
comprise two filler sequences, the first filler sequence may be
located between the 5' ITR and exon region, and the second filler
sequence may be located between the polyadenylation signal sequence
region and 3' ITR. In one embodiment, a viral genome may comprise
two filler sequences, the first filler sequence may be located
between the 5' ITR and exon region, and the second filler sequence
may be located between the MCS region and exon region. In one
embodiment, a viral genome may comprise two filler sequences, the
first filler sequence may be located between the 5' ITR and exon
region, and the second filler sequence may be located between the
MCS region and 3' ITR. In one embodiment, a viral genome may
comprise two filler sequences, the first filler sequence may be
located between the 5' ITR and exon region, and the second filler
sequence may be located between the exon region and 3' ITR.
[0190] In one embodiment, a viral genome may comprise two filler
sequences, the first filler sequence may be located between the
promoter region and payload region, and the second filler sequence
may be located between the payload region and intron region. In one
embodiment, a viral genome may comprise two filler sequences, the
first filler sequence may be located between the promoter region
and payload region, and the second filler sequence may be located
between the payload region and enhancer region. In one embodiment,
a viral genome may comprise two filler sequences, the first filler
sequence may be located between the promoter region and payload
region, and the second filler sequence may be located between the
payload region and polyadenylation signal sequence region. In one
embodiment, a viral genome may comprise two filler sequences, the
first filler sequence may be located between the promoter region
and payload region, and the second filler sequence may be located
between the payload region and MCS region. In one embodiment, a
viral genome may comprise two filler sequences, the first filler
sequence may be located between the promoter region and payload
region, and the second filler sequence may be located between the
payload region and exon region. In one embodiment, a viral genome
may comprise two filler sequences, the first filler sequence may be
located between the promoter region and payload region, and the
second filler sequence may be located between the payload region
and 3' ITR. In one embodiment, a viral genome may comprise two
filler sequences, the first filler sequence may be located between
the promoter region and payload region, and the second filler
sequence may be located between the intron region and enhancer
region. In one embodiment, a viral genome may comprise two filler
sequences, the first filler sequence may be located between the
promoter region and payload region, and the second filler sequence
may be located between the intron region and polyadenylation signal
sequence region. In one embodiment, a viral genome may comprise two
filler sequences, the first filler sequence may be located between
the promoter region and payload region, and the second filler
sequence may be located between the intron region and MCS region.
In one embodiment, a viral genome may comprise two filler
sequences, the first filler sequence may be located between the
promoter region and payload region, and the second filler sequence
may be located between the intron region and exon region. In one
embodiment, a viral genome may comprise two filler sequences, the
first filler sequence may be located between the promoter region
and payload region, and the second filler sequence may be located
between the intron region and 3' ITR. In one embodiment, a viral
genome may comprise two filler sequences, the first filler sequence
may be located between the promoter region and payload region, and
the second filler sequence may be located between the enhancer
region and polyadenylation signal sequence region. In one
embodiment, a viral genome may comprise two filler sequences, the
first filler sequence may be located between the promoter region
and payload region, and the second filler sequence may be located
between the enhancer region and MCS region. In one embodiment, a
viral genome may comprise two filler sequences, the first filler
sequence may be located between the promoter region and payload
region, and the second filler sequence may be located between the
enhancer region and exon region. In one embodiment, a viral genome
may comprise two filler sequences, the first filler sequence may be
located between the promoter region and payload region, and the
second filler sequence may be located between the enhancer region
and 3' ITR. In one embodiment, a viral genome may comprise two
filler sequences, the first filler sequence may be located between
the promoter region and payload region, and the second filler
sequence may be located between the polyadenylation signal sequence
region and MCS region. In one embodiment, a viral genome may
comprise two filler sequences, the first filler sequence may be
located between the promoter region and payload region, and the
second filler sequence may be located between the polyadenylation
signal sequence region and exon region. In one embodiment, a viral
genome may comprise two filler sequences, the first filler sequence
may be located between the promoter region and payload region, and
the second filler sequence may be located between the
polyadenylation signal sequence region and 3' ITR. In one
embodiment, a viral genome may comprise two filler sequences, the
first filler sequence may be located between the promoter region
and payload region, and the second filler sequence may be located
between the MCS region and exon region. In one embodiment, a viral
genome may comprise two filler sequences, the first filler sequence
may be located between the promoter region and payload region, and
the second filler sequence may be located between the MCS region
and 3' ITR. In one embodiment, a viral genome may comprise two
filler sequences, the first filler sequence may be located between
the promoter region and payload region, and the second filler
sequence may be located between the exon region and 3' ITR.
[0191] In one embodiment, a viral genome may comprise two filler
sequences, the first filler sequence may be located between the
promoter region and intron region, and the second filler sequence
may be located between the payload region and intron region. In one
embodiment, a viral genome may comprise two filler sequences, the
first filler sequence may be located between the promoter region
and intron region, and the second filler sequence may be located
between the payload region and enhancer region. In one embodiment,
a viral genome may comprise two filler sequences, the first filler
sequence may be located between the promoter region and intron
region, and the second filler sequence may be located between the
payload region and polyadenylation signal sequence region. In one
embodiment, a viral genome may comprise two filler sequences, the
first filler sequence may be located between the promoter region
and intron region, and the second filler sequence may be located
between the payload region and MCS region. In one embodiment, a
viral genome may comprise two filler sequences, the first filler
sequence may be located between the promoter region and intron
region, and the second filler sequence may be located between the
payload region and exon region. In one embodiment, a viral genome
may comprise two filler sequences, the first filler sequence may be
located between the promoter region and intron region, and the
second filler sequence may be located between the payload region
and 3' ITR. In one embodiment, a viral genome may comprise two
filler sequences, the first filler sequence may be located between
the promoter region and intron region, and the second filler
sequence may be located between the intron region and enhancer
region. In one embodiment, a viral genome may comprise two filler
sequences, the first filler sequence may be located between the
promoter region and intron region, and the second filler sequence
may be located between the intron region and polyadenylation signal
sequence region. In one embodiment, a viral genome may comprise two
filler sequences, the first filler sequence may be located between
the promoter region and intron region, and the second filler
sequence may be located between the intron region and MCS region.
In one embodiment, a viral genome may comprise two filler
sequences, the first filler sequence may be located between the
promoter region and intron region, and the second filler sequence
may be located between the intron region and exon region. In one
embodiment, a viral genome may comprise two filler sequences, the
first filler sequence may be located between the promoter region
and intron region, and the second filler sequence may be located
between the intron region and 3' ITR. In one embodiment, a viral
genome may comprise two filler sequences, the first filler sequence
may be located between the promoter region and intron region, and
the second filler sequence may be located between the enhancer
region and polyadenylation signal sequence region. In one
embodiment, a viral genome may comprise two filler sequences, the
first filler sequence may be located between the promoter region
and intron region, and the second filler sequence may be located
between the enhancer region and MCS region. In one embodiment, a
viral genome may comprise two filler sequences, the first filler
sequence may be located between the promoter region and intron
region, and the second filler sequence may be located between the
enhancer region and exon region. In one embodiment, a viral genome
may comprise two filler sequences, the first filler sequence may be
located between the promoter region and intron region, and the
second filler sequence may be located between the enhancer region
and 3' ITR. In one embodiment, a viral genome may comprise two
filler sequences, the first filler sequence may be located between
the promoter region and intron region, and the second filler
sequence may be located between the polyadenylation signal sequence
region and MCS region. In one embodiment, a viral genome may
comprise two filler sequences, the first filler sequence may be
located between the promoter region and intron region, and the
second filler sequence may be located between the polyadenylation
signal sequence region and exon region. In one embodiment, a viral
genome may comprise two filler sequences, the first filler sequence
may be located between the promoter region and intron region, and
the second filler sequence may be located between the
polyadenylation signal sequence region and 3' ITR. In one
embodiment, a viral genome may comprise two filler sequences, the
first filler sequence may be located between the promoter region
and intron region, and the second filler sequence may be located
between the MCS region and exon region. In one embodiment, a viral
genome may comprise two filler sequences, the first filler sequence
may be located between the promoter region and intron region, and
the second filler sequence may be located between the MCS region
and 3' ITR. In one embodiment, a viral genome may comprise two
filler sequences, the first filler sequence may be located between
the promoter region and intron region, and the second filler
sequence may be located between the exon region and 3' ITR.
[0192] In one embodiment, a viral genome may comprise two filler
sequences, the first filler sequence may be located between the
promoter region and enhancer region, and the second filler sequence
may be located between the payload region and intron region. In one
embodiment, a viral genome may comprise two filler sequences, the
first filler sequence may be located between the promoter region
and enhancer region, and the second filler sequence may be located
between the payload region and enhancer region. In one embodiment,
a viral genome may comprise two filler sequences, the first filler
sequence may be located between the promoter region and enhancer
region, and the second filler sequence may be located between the
payload region and polyadenylation signal sequence region. In one
embodiment, a viral genome may comprise two filler sequences, the
first filler sequence may be located between the promoter region
and enhancer region, and the second filler sequence may be located
between the payload region and MCS region. In one embodiment, a
viral genome may comprise two filler sequences, the first filler
sequence may be located between the promoter region and enhancer
region, and the second filler sequence may be located between the
payload region and exon region. In one embodiment, a viral genome
may comprise two filler sequences, the first filler sequence may be
located between the promoter region and enhancer region, and the
second filler sequence may be located between the payload region
and 3' ITR. In one embodiment, a viral genome may comprise two
filler sequences, the first filler sequence may be located between
the promoter region and enhancer region, and the second filler
sequence may be located between the intron region and enhancer
region. In one embodiment, a viral genome may comprise two filler
sequences, the first filler sequence may be located between the
promoter region and enhancer region, and the second filler sequence
may be located between the intron region and polyadenylation signal
sequence region. In one embodiment, a viral genome may comprise two
filler sequences, the first filler sequence may be located between
the promoter region and enhancer region, and the second filler
sequence may be located between the intron region and MCS region.
In one embodiment, a viral genome may comprise two filler
sequences, the first filler sequence may be located between the
promoter region and enhancer region, and the second filler sequence
may be located between the intron region and exon region. In one
embodiment, a viral genome may comprise two filler sequences, the
first filler sequence may be located between the promoter region
and enhancer region, and the second filler sequence may be located
between the intron region and 3' ITR. In one embodiment, a viral
genome may comprise two filler sequences, the first filler sequence
may be located between the promoter region and enhancer region, and
the second filler sequence may be located between the enhancer
region and polyadenylation signal sequence region. In one
embodiment, a viral genome may comprise two filler sequences, the
first filler sequence may be located between the promoter region
and enhancer region, and the second filler sequence may be located
between the enhancer region and MCS region. In one embodiment, a
viral genome may comprise two filler sequences, the first filler
sequence may be located between the promoter region and enhancer
region, and the second filler sequence may be located between the
enhancer region and exon region. In one embodiment, a viral genome
may comprise two filler sequences, the first filler sequence may be
located between the promoter region and enhancer region, and the
second filler sequence may be located between the enhancer region
and 3' ITR. In one embodiment, a viral genome may comprise two
filler sequences, the first filler sequence may be located between
the promoter region and enhancer region, and the second filler
sequence may be located between the polyadenylation signal sequence
region and MCS region. In one embodiment, a viral genome may
comprise two filler sequences, the first filler sequence may be
located between the promoter region and enhancer region, and the
second filler sequence may be located between the polyadenylation
signal sequence region and exon region. In one embodiment, a viral
genome may comprise two filler sequences, the first filler sequence
may be located between the promoter region and enhancer region, and
the second filler sequence may be located between the
polyadenylation signal sequence region and 3' ITR. In one
embodiment, a viral genome may comprise two filler sequences, the
first filler sequence may be located between the promoter region
and enhancer region, and the second filler sequence may be located
between the MCS region and exon region. In one embodiment, a viral
genome may comprise two filler sequences, the first filler sequence
may be located between the promoter region and enhancer region, and
the second filler sequence may be located between the MCS region
and 3' ITR. In one embodiment, a viral genome may comprise two
filler sequences, the first filler sequence may be located between
the promoter region and enhancer region, and the second filler
sequence may be located between the exon region and 3' ITR.
[0193] In one embodiment, a viral genome may comprise two filler
sequences, the first filler sequence may be located between the
promoter region and polyadenylation signal sequence region, and the
second filler sequence may be located between the payload region
and intron region. In one embodiment, a viral genome may comprise
two filler sequences, the first filler sequence may be located
between the promoter region and polyadenylation signal sequence
region, and the second filler sequence may be located between the
payload region and enhancer region. In one embodiment, a viral
genome may comprise two filler sequences, the first filler sequence
may be located between the promoter region and polyadenylation
signal sequence region, and the second filler sequence may be
located between the payload region and polyadenylation signal
sequence region. In one embodiment, a viral genome may comprise two
filler sequences, the first filler sequence may be located between
the promoter region and polyadenylation signal sequence region, and
the second filler sequence may be located between the payload
region and MCS region. In one embodiment, a viral genome may
comprise two filler sequences, the first filler sequence may be
located between the promoter region and polyadenylation signal
sequence region, and the second filler sequence may be located
between the payload region and exon region. In one embodiment, a
viral genome may comprise two filler sequences, the first filler
sequence may be located between the promoter region and
polyadenylation signal sequence region, and the second filler
sequence may be located between the payload region and 3' ITR. In
one embodiment, a viral genome may comprise two filler sequences,
the first filler sequence may be located between the promoter
region and polyadenylation signal sequence region, and the second
filler sequence may be located between the intron region and
enhancer region. In one embodiment, a viral genome may comprise two
filler sequences, the first filler sequence may be located between
the promoter region and polyadenylation signal sequence region, and
the second filler sequence may be located between the intron region
and polyadenylation signal sequence region. In one embodiment, a
viral genome may comprise two filler sequences, the first filler
sequence may be located between the promoter region and
polyadenylation signal sequence region, and the second filler
sequence may be located between the intron region and MCS region.
In one embodiment, a viral genome may comprise two filler
sequences, the first filler sequence may be located between the
promoter region and polyadenylation signal sequence region, and the
second filler sequence may be located between the intron region and
exon region. In one embodiment, a viral genome may comprise two
filler sequences, the first filler sequence may be located between
the promoter region and polyadenylation signal sequence region, and
the second filler sequence may be located between the intron region
and 3' ITR. In one embodiment, a viral genome may comprise two
filler sequences, the first filler sequence may be located between
the promoter region and polyadenylation signal sequence region, and
the second filler sequence may be located between the enhancer
region and polyadenylation signal sequence region. In one
embodiment, a viral genome may comprise two filler sequences, the
first filler sequence may be located between the promoter region
and polyadenylation signal sequence region, and the second filler
sequence may be located between the enhancer region and MCS region.
In one embodiment, a viral genome may comprise two filler
sequences, the first filler sequence may be located between the
promoter region and polyadenylation signal sequence region, and the
second filler sequence may be located between the enhancer region
and exon region. In one embodiment, a viral genome may comprise two
filler sequences, the first filler sequence may be located between
the promoter region and polyadenylation signal sequence region, and
the second filler sequence may be located between the enhancer
region and 3' ITR. In one embodiment, a viral genome may comprise
two filler sequences, the first filler sequence may be located
between the promoter region and polyadenylation signal sequence
region, and the second filler sequence may be located between the
polyadenylation signal sequence region and MCS region. In one
embodiment, a viral genome may comprise two filler sequences, the
first filler sequence may be located between the promoter region
and polyadenylation signal sequence region, and the second filler
sequence may be located between the polyadenylation signal sequence
region and exon region. In one embodiment, a viral genome may
comprise two filler sequences, the first filler sequence may be
located between the promoter region and polyadenylation signal
sequence region, and the second filler sequence may be located
between the polyadenylation signal sequence region and 3' ITR. In
one embodiment, a viral genome may comprise two filler sequences,
the first filler sequence may be located between the promoter
region and polyadenylation signal sequence region, and the second
filler sequence may be located between the MCS region and exon
region. In one embodiment, a viral genome may comprise two filler
sequences, the first filler sequence may be located between the
promoter region and polyadenylation signal sequence region, and the
second filler sequence may be located between the MCS region and 3'
ITR. In one embodiment, a viral genome may comprise two filler
sequences, the first filler sequence may be located between the
promoter region and polyadenylation signal sequence region, and the
second filler sequence may be located between the exon region and
3' ITR.
[0194] In one embodiment, a viral genome may comprise two filler
sequences, the first filler sequence may be located between the
promoter region and exon region, and the second filler sequence may
be located between the payload region and intron region. In one
embodiment, a viral genome may comprise two filler sequences, the
first filler sequence may be located between the promoter region
and exon region, and the second filler sequence may be located
between the payload region and enhancer region. In one embodiment,
a viral genome may comprise two filler sequences, the first filler
sequence may be located between the promoter region and exon
region, and the second filler sequence may be located between the
payload region and polyadenylation signal sequence region. In one
embodiment, a viral genome may comprise two filler sequences, the
first filler sequence may be located between the promoter region
and exon region, and the second filler sequence may be located
between the payload region and MCS region. In one embodiment, a
viral genome may comprise two filler sequences, the first filler
sequence may be located between the promoter region and exon
region, and the second filler sequence may be located between the
payload region and exon region. In one embodiment, a viral genome
may comprise two filler sequences, the first filler sequence may be
located between the promoter region and exon region, and the second
filler sequence may be located between the payload region and 3'
ITR. In one embodiment, a viral genome may comprise two filler
sequences, the first filler sequence may be located between the
promoter region and exon region, and the second filler sequence may
be located between the intron region and enhancer region. In one
embodiment, a viral genome may comprise two filler sequences, the
first filler sequence may be located between the promoter region
and exon region, and the second filler sequence may be located
between the intron region and polyadenylation signal sequence
region. In one embodiment, a viral genome may comprise two filler
sequences, the first filler sequence may be located between the
promoter region and exon region, and the second filler sequence may
be located between the intron region and MCS region. In one
embodiment, a viral genome may comprise two filler sequences, the
first filler sequence may be located between the promoter region
and exon region, and the second filler sequence may be located
between the intron region and exon region. In one embodiment, a
viral genome may comprise two filler sequences, the first filler
sequence may be located between the promoter region and exon
region, and the second filler sequence may be located between the
intron region and 3' ITR. In one embodiment, a viral genome may
comprise two filler sequences, the first filler sequence may be
located between the promoter region and exon region, and the second
filler sequence may be located between the enhancer region and
polyadenylation signal sequence region. In one embodiment, a viral
genome may comprise two filler sequences, the first filler sequence
may be located between the promoter region and exon region, and the
second filler sequence may be located between the enhancer region
and MCS region. In one embodiment, a viral genome may comprise two
filler sequences, the first filler sequence may be located between
the promoter region and exon region, and the second filler sequence
may be located between the enhancer region and exon region. In one
embodiment, a viral genome may comprise two filler sequences, the
first filler sequence may be located between the promoter region
and exon region, and the second filler sequence may be located
between the enhancer region and 3' ITR. In one embodiment, a viral
genome may comprise two filler sequences, the first filler sequence
may be located between the promoter region and exon region, and the
second filler sequence may be located between the polyadenylation
signal sequence region and MCS region. In one embodiment, a viral
genome may comprise two filler sequences, the first filler sequence
may be located between the promoter region and exon region, and the
second filler sequence may be located between the polyadenylation
signal sequence region and exon region. In one embodiment, a viral
genome may comprise two filler sequences, the first filler sequence
may be located between the promoter region and exon region, and the
second filler sequence may be located between the polyadenylation
signal sequence region and 3' ITR. In one embodiment, a viral
genome may comprise two filler sequences, the first filler sequence
may be located between the promoter region and exon region, and the
second filler sequence may be located between the MCS region and
exon region. In one embodiment, a viral genome may comprise two
filler sequences, the first filler sequence may be located between
the promoter region and exon region, and the second filler sequence
may be located between the MCS region and 3' ITR. In one
embodiment, a viral genome may comprise two filler sequences, the
first filler sequence may be located between the promoter region
and exon region, and the second filler sequence may be located
between the exon region and 3' ITR.
[0195] In one embodiment, a viral genome may comprise two filler
sequences, the first filler sequence may be located between the
promoter region and MCS region, and the second filler sequence may
be located between the payload region and intron region. In one
embodiment, a viral genome may comprise two filler sequences, the
first filler sequence may be located between the promoter region
and MCS region, and the second filler sequence may be located
between the payload region and enhancer region. In one embodiment,
a viral genome may comprise two filler sequences, the first filler
sequence may be located between the promoter region and MCS region,
and the second filler sequence may be located between the payload
region and polyadenylation signal sequence region. In one
embodiment, a viral genome may comprise two filler sequences, the
first filler sequence may be located between the promoter region
and MCS region, and the second filler sequence may be located
between the payload region and MCS region. In one embodiment, a
viral genome may comprise two filler sequences, the first filler
sequence may be located between the promoter region and MCS region,
and the second filler sequence may be located between the payload
region and exon region. In one embodiment, a viral genome may
comprise two filler sequences, the first filler sequence may be
located between the promoter region and MCS region, and the second
filler sequence may be located between the payload region and 3'
ITR. In one embodiment, a viral genome may comprise two filler
sequences, the first filler sequence may be located between the
promoter region and MCS region, and the second filler sequence may
be located between the intron region and enhancer region. In one
embodiment, a viral genome may comprise two filler sequences, the
first filler sequence may be located between the promoter region
and MCS region, and the second filler sequence may be located
between the intron region and polyadenylation signal sequence
region. In one embodiment, a viral genome may comprise two filler
sequences, the first filler sequence may be located between the
promoter region and MCS region, and the second filler sequence may
be located between the intron region and MCS region. In one
embodiment, a viral genome may comprise two filler sequences, the
first filler sequence may be located between the promoter region
and MCS region, and the second filler sequence may be located
between the intron region and exon region. In one embodiment, a
viral genome may comprise two filler sequences, the first filler
sequence may be located between the promoter region and MCS region,
and the second filler sequence may be located between the intron
region and 3' ITR. In one embodiment, a viral genome may comprise
two filler sequences, the first filler sequence may be located
between the promoter region and MCS region, and the second filler
sequence may be located between the enhancer region and
polyadenylation signal sequence region. In one embodiment, a viral
genome may comprise two filler sequences, the first filler sequence
may be located between the promoter region and MCS region, and the
second filler sequence may be located between the enhancer region
and MCS region. In one embodiment, a viral genome may comprise two
filler sequences, the first filler sequence may be located between
the promoter region and MCS region, and the second filler sequence
may be located between the enhancer region and exon region. In one
embodiment, a viral genome may comprise two filler sequences, the
first filler sequence may be located between the promoter region
and MCS region, and the second filler sequence may be located
between the enhancer region and 3' ITR. In one embodiment, a viral
genome may comprise two filler sequences, the first filler sequence
may be located between the promoter region and MCS region, and the
second filler sequence may be located between the polyadenylation
signal sequence region and MCS region. In one embodiment, a viral
genome may comprise two filler sequences, the first filler sequence
may be located between the promoter region and MCS region, and the
second filler sequence may be located between the polyadenylation
signal sequence region and exon region. In one embodiment, a viral
genome may comprise two filler sequences, the first filler sequence
may be located between the promoter region and MCS region, and the
second filler sequence may be located between the polyadenylation
signal sequence region and 3' ITR. In one embodiment, a viral
genome may comprise two filler sequences, the first filler sequence
may be located between the promoter region and MCS region, and the
second filler sequence may be located between the MCS region and
exon region. In one embodiment, a viral genome may comprise two
filler sequences, the first filler sequence may be located between
the promoter region and MCS region, and the second filler sequence
may be located between the MCS region and 3' ITR. In one
embodiment, a viral genome may comprise two filler sequences, the
first filler sequence may be located between the promoter region
and MCS region, and the second filler sequence may be located
between the exon region and 3' ITR.
[0196] In one embodiment, a viral genome may comprise two filler
sequences, the first filler sequence may be located between the
promoter region and 3'ITR, and the second filler sequence may be
located between the payload region and intron region. In one
embodiment, a viral genome may comprise two filler sequences, the
first filler sequence may be located between the promoter region
and 3'ITR, and the second filler sequence may be located between
the payload region and enhancer region. In one embodiment, a viral
genome may comprise two filler sequences, the first filler sequence
may be located between the promoter region and 3'ITR, and the
second filler sequence may be located between the payload region
and polyadenylation signal sequence region. In one embodiment, a
viral genome may comprise two filler sequences, the first filler
sequence may be located between the promoter region and 3'ITR, and
the second filler sequence may be located between the payload
region and MCS region. In one embodiment, a viral genome may
comprise two filler sequences, the first filler sequence may be
located between the promoter region and 3'ITR, and the second
filler sequence may be located between the payload region and exon
region. In one embodiment, a viral genome may comprise two filler
sequences, the first filler sequence may be located between the
promoter region and 3'ITR, and the second filler sequence may be
located between the payload region and 3' ITR. In one embodiment, a
viral genome may comprise two filler sequences, the first filler
sequence may be located between the promoter region and 3'ITR, and
the second filler sequence may be located between the intron region
and enhancer region. In one embodiment, a viral genome may comprise
two filler sequences, the first filler sequence may be located
between the promoter region and 3'ITR, and the second filler
sequence may be located between the intron region and
polyadenylation signal sequence region. In one embodiment, a viral
genome may comprise two filler sequences, the first filler sequence
may be located between the promoter region and 3'ITR, and the
second filler sequence may be located between the intron region and
MCS region. In one embodiment, a viral genome may comprise two
filler sequences, the first filler sequence may be located between
the promoter region and 3'ITR, and the second filler sequence may
be located between the intron region and exon region. In one
embodiment, a viral genome may comprise two filler sequences, the
first filler sequence may be located between the promoter region
and 3'ITR, and the second filler sequence may be located between
the intron region and 3' ITR. In one embodiment, a viral genome may
comprise two filler sequences, the first filler sequence may be
located between the promoter region and 3'ITR, and the second
filler sequence may be located between the enhancer region and
polyadenylation signal sequence region. In one embodiment, a viral
genome may comprise two filler sequences, the first filler sequence
may be located between the promoter region and 3'ITR, and the
second filler sequence may be located between the enhancer region
and MCS region. In one embodiment, a viral genome may comprise two
filler sequences, the first filler sequence may be located between
the promoter region and 3'ITR, and the second filler sequence may
be located between the enhancer region and exon region. In one
embodiment, a viral genome may comprise two filler sequences, the
first filler sequence may be located between the promoter region
and 3'ITR, and the second filler sequence may be located between
the enhancer region and 3' ITR. In one embodiment, a viral genome
may comprise two filler sequences, the first filler sequence may be
located between the promoter region and 3'ITR, and the second
filler sequence may be located between the polyadenylation signal
sequence region and MCS region. In one embodiment, a viral genome
may comprise two filler sequences, the first filler sequence may be
located between the promoter region and 3'ITR, and the second
filler sequence may be located between the polyadenylation signal
sequence region and exon region. In one embodiment, a viral genome
may comprise two filler sequences, the first filler sequence may be
located between the promoter region and 3'ITR, and the second
filler sequence may be located between the polyadenylation signal
sequence region and 3' ITR. In one embodiment, a viral genome may
comprise two filler sequences, the first filler sequence may be
located between the promoter region and 3'ITR, and the second
filler sequence may be located between the MCS region and exon
region. In one embodiment, a viral genome may comprise two filler
sequences, the first filler sequence may be located between the
promoter region and 3'ITR, and the second filler sequence may be
located between the MCS region and 3' ITR. In one embodiment, a
viral genome may comprise two filler sequences, the first filler
sequence may be located between the promoter region and 3'ITR, and
the second filler sequence may be located between the exon region
and 3' ITR.
[0197] In one embodiment, a viral genome may comprise two filler
sequences, the first filler sequence may be located between the
payload region and intron region, and the second filler sequence
may be located between the intron region and enhancer region. In
one embodiment, a viral genome may comprise two filler sequences,
the first filler sequence may be located between the payload region
and intron region, and the second filler sequence may be located
between the intron region and polyadenylation signal sequence
region. In one embodiment, a viral genome may comprise two filler
sequences, the first filler sequence may be located between the
payload region and intron region, and the second filler sequence
may be located between the intron region and MCS region. In one
embodiment, a viral genome may comprise two filler sequences, the
first filler sequence may be located between the payload region and
intron region, and the second filler sequence may be located
between the intron region and exon region. In one embodiment, a
viral genome may comprise two filler sequences, the first filler
sequence may be located between the payload region and intron
region, and the second filler sequence may be located between the
intron region and 3' ITR. In one embodiment, a viral genome may
comprise two filler sequences, the first filler sequence may be
located between the payload region and intron region, and the
second filler sequence may be located between the enhancer region
and polyadenylation signal sequence region. In one embodiment, a
viral genome may comprise two filler sequences, the first filler
sequence may be located between the payload region and intron
region, and the second filler sequence may be located between the
enhancer region and MCS region. In one embodiment, a viral genome
may comprise two filler sequences, the first filler sequence may be
located between the payload region and intron region, and the
second filler sequence may be located between the enhancer region
and exon region. In one embodiment, a viral genome may comprise two
filler sequences, the first filler sequence may be located between
the payload region and intron region, and the second filler
sequence may be located between the enhancer region and 3' ITR. In
one embodiment, a viral genome may comprise two filler sequences,
the first filler sequence may be located between the payload region
and intron region, and the second filler sequence may be located
between the polyadenylation signal sequence region and MCS region.
In one embodiment, a viral genome may comprise two filler
sequences, the first filler sequence may be located between the
payload region and intron region, and the second filler sequence
may be located between the polyadenylation signal sequence region
and exon region. In one embodiment, a viral genome may comprise two
filler sequences, the first filler sequence may be located between
the payload region and intron region, and the second filler
sequence may be located between the polyadenylation signal sequence
region and 3' ITR. In one embodiment, a viral genome may comprise
two filler sequences, the first filler sequence may be located
between the payload region and intron region, and the second filler
sequence may be located between the MCS region and exon region. In
one embodiment, a viral genome may comprise two filler sequences,
the first filler sequence may be located between the payload region
and intron region, and the second filler sequence may be located
between the MCS region and 3' ITR. In one embodiment, a viral
genome may comprise two filler sequences, the first filler sequence
may be located between the payload region and intron region, and
the second filler sequence may be located between the exon region
and 3' ITR.
[0198] In one embodiment, a viral genome may comprise two filler
sequences, the first filler sequence may be located between the
payload region and enhancer region, and the second filler sequence
may be located between the intron region and enhancer region. In
one embodiment, a viral genome may comprise two filler sequences,
the first filler sequence may be located between the payload region
and enhancer region, and the second filler sequence may be located
between the intron region and polyadenylation signal sequence
region. In one embodiment, a viral genome may comprise two filler
sequences, the first filler sequence may be located between the
payload region and enhancer region, and the second filler sequence
may be located between the intron region and MCS region. In one
embodiment, a viral genome may comprise two filler sequences, the
first filler sequence may be located between the payload region and
enhancer region, and the second filler sequence may be located
between the intron region and exon region. In one embodiment, a
viral genome may comprise two filler sequences, the first filler
sequence may be located between the payload region and enhancer
region, and the second filler sequence may be located between the
intron region and 3' ITR. In one embodiment, a viral genome may
comprise two filler sequences, the first filler sequence may be
located between the payload region and enhancer region, and the
second filler sequence may be located between the enhancer region
and polyadenylation signal sequence region. In one embodiment, a
viral genome may comprise two filler sequences, the first filler
sequence may be located between the payload region and enhancer
region, and the second filler sequence may be located between the
enhancer region and MCS region. In one embodiment, a viral genome
may comprise two filler sequences, the first filler sequence may be
located between the payload region and enhancer region, and the
second filler sequence may be located between the enhancer region
and exon region. In one embodiment, a viral genome may comprise two
filler sequences, the first filler sequence may be located between
the payload region and enhancer region, and the second filler
sequence may be located between the enhancer region and 3' ITR. In
one embodiment, a viral genome may comprise two filler sequences,
the first filler sequence may be located between the payload region
and enhancer region, and the second filler sequence may be located
between the polyadenylation signal sequence region and MCS region.
In one embodiment, a viral genome may comprise two filler
sequences, the first filler sequence may be located between the
payload region and enhancer region, and the second filler sequence
may be located between the polyadenylation signal sequence region
and exon region. In one embodiment, a viral genome may comprise two
filler sequences, the first filler sequence may be located between
the payload region and enhancer region, and the second filler
sequence may be located between the polyadenylation signal sequence
region and 3' ITR. In one embodiment, a viral genome may comprise
two filler sequences, the first filler sequence may be located
between the payload region and enhancer region, and the second
filler sequence may be located between the MCS region and exon
region. In one embodiment, a viral genome may comprise two filler
sequences, the first filler sequence may be located between the
payload region and enhancer region, and the second filler sequence
may be located between the MCS region and 3' ITR. In one
embodiment, a viral genome may comprise two filler sequences, the
first filler sequence may be located between the payload region and
enhancer region, and the second filler sequence may be located
between the exon region and 3' ITR.
[0199] In one embodiment, a viral genome may comprise two filler
sequences, the first filler sequence may be located between the
payload region and polyadenylation signal sequence region, and the
second filler sequence may be located between the intron region and
enhancer region. In one embodiment, a viral genome may comprise two
filler sequences, the first filler sequence may be located between
the payload region and polyadenylation signal sequence region, and
the second filler sequence may be located between the intron region
and polyadenylation signal sequence region. In one embodiment, a
viral genome may comprise two filler sequences, the first filler
sequence may be located between the payload region and
polyadenylation signal sequence region, and the second filler
sequence may be located between the intron region and MCS region.
In one embodiment, a viral genome may comprise two filler
sequences, the first filler sequence may be located between the
payload region and polyadenylation signal sequence region, and the
second filler sequence may be located between the intron region and
exon region. In one embodiment, a viral genome may comprise two
filler sequences, the first filler sequence may be located between
the payload region and polyadenylation signal sequence region, and
the second filler sequence may be located between the intron region
and 3' ITR. In one embodiment, a viral genome may comprise two
filler sequences, the first filler sequence may be located between
the payload region and polyadenylation signal sequence region, and
the second filler sequence may be located between the enhancer
region and polyadenylation signal sequence region. In one
embodiment, a viral genome may comprise two filler sequences, the
first filler sequence may be located between the payload region and
polyadenylation signal sequence region, and the second filler
sequence may be located between the enhancer region and MCS region.
In one embodiment, a viral genome may comprise two filler
sequences, the first filler sequence may be located between the
payload region and polyadenylation signal sequence region, and the
second filler sequence may be located between the enhancer region
and exon region. In one embodiment, a viral genome may comprise two
filler sequences, the first filler sequence may be located between
the payload region and polyadenylation signal sequence region, and
the second filler sequence may be located between the enhancer
region and 3' ITR. In one embodiment, a viral genome may comprise
two filler sequences, the first filler sequence may be located
between the payload region and polyadenylation signal sequence
region, and the second filler sequence may be located between the
polyadenylation signal sequence region and MCS region. In one
embodiment, a viral genome may comprise two filler sequences, the
first filler sequence may be located between the payload region and
polyadenylation signal sequence region, and the second filler
sequence may be located between the polyadenylation signal sequence
region and exon region. In one embodiment, a viral genome may
comprise two filler sequences, the first filler sequence may be
located between the payload region and polyadenylation signal
sequence region, and the second filler sequence may be located
between the polyadenylation signal sequence region and 3' ITR. In
one embodiment, a viral genome may comprise two filler sequences,
the first filler sequence may be located between the payload region
and polyadenylation signal sequence region, and the second filler
sequence may be located between the MCS region and exon region. In
one embodiment, a viral genome may comprise two filler sequences,
the first filler sequence may be located between the payload region
and polyadenylation signal sequence region, and the second filler
sequence may be located between the MCS region and 3' ITR. In one
embodiment, a viral genome may comprise two filler sequences, the
first filler sequence may be located between the payload region and
polyadenylation signal sequence region, and the second filler
sequence may be located between the exon region and 3' ITR.
[0200] In one embodiment, a viral genome may comprise two filler
sequences, the first filler sequence may be located between the
payload region and MCS region, and the second filler sequence may
be located between the intron region and enhancer region. In one
embodiment, a viral genome may comprise two filler sequences, the
first filler sequence may be located between the payload region and
MCS region, and the second filler sequence may be located between
the intron region and polyadenylation signal sequence region. In
one embodiment, a viral genome may comprise two filler sequences,
the first filler sequence may be located between the payload region
and MCS region, and the second filler sequence may be located
between the intron region and MCS region. In one embodiment, a
viral genome may comprise two filler sequences, the first filler
sequence may be located between the payload region and MCS region,
and the second filler sequence may be located between the intron
region and exon region. In one embodiment, a viral genome may
comprise two filler sequences, the first filler sequence may be
located between the payload region and MCS region, and the second
filler sequence may be located between the intron region and 3'
ITR. In one embodiment, a viral genome may comprise two filler
sequences, the first filler sequence may be located between the
payload region and MCS region, and the second filler sequence may
be located between the enhancer region and polyadenylation signal
sequence region. In one embodiment, a viral genome may comprise two
filler sequences, the first filler sequence may be located between
the payload region and MCS region, and the second filler sequence
may be located between the enhancer region and MCS region. In one
embodiment, a viral genome may comprise two filler sequences, the
first filler sequence may be located between the payload region and
MCS region, and the second filler sequence may be located between
the enhancer region and exon region. In one embodiment, a viral
genome may comprise two filler sequences, the first filler sequence
may be located between the payload region and MCS region, and the
second filler sequence may be located between the enhancer region
and 3' ITR. In one embodiment, a viral genome may comprise two
filler sequences, the first filler sequence may be located between
the payload region and MCS region, and the second filler sequence
may be located between the polyadenylation signal sequence region
and MCS region. In one embodiment, a viral genome may comprise two
filler sequences, the first filler sequence may be located between
the payload region and MCS region, and the second filler sequence
may be located between the polyadenylation signal sequence region
and exon region. In one embodiment, a viral genome may comprise two
filler sequences, the first filler sequence may be located between
the payload region and MCS region, and the second filler sequence
may be located between the polyadenylation signal sequence region
and 3' ITR. In one embodiment, a viral genome may comprise two
filler sequences, the first filler sequence may be located between
the payload region and MCS region, and the second filler sequence
may be located between the MCS region and exon region. In one
embodiment, a viral genome may comprise two filler sequences, the
first filler sequence may be located between the payload region and
MCS region, and the second filler sequence may be located between
the MCS region and 3' ITR. In one embodiment, a viral genome may
comprise two filler sequences, the first filler sequence may be
located between the payload region and MCS region, and the second
filler sequence may be located between the exon region and 3'
ITR.
[0201] In one embodiment, a viral genome may comprise two filler
sequences, the first filler sequence may be located between the
payload region and exon region, and the second filler sequence may
be located between the intron region and enhancer region. In one
embodiment, a viral genome may comprise two filler sequences, the
first filler sequence may be located between the payload region and
exon region, and the second filler sequence may be located between
the intron region and polyadenylation signal sequence region. In
one embodiment, a viral genome may comprise two filler sequences,
the first filler sequence may be located between the payload region
and exon region, and the second filler sequence may be located
between the intron region and MCS region. In one embodiment, a
viral genome may comprise two filler sequences, the first filler
sequence may be located between the payload region and exon region,
and the second filler sequence may be located between the intron
region and exon region. In one embodiment, a viral genome may
comprise two filler sequences, the first filler sequence may be
located between the payload region and exon region, and the second
filler sequence may be located between the intron region and 3'
ITR. In one embodiment, a viral genome may comprise two filler
sequences, the first filler sequence may be located between the
payload region and exon region, and the second filler sequence may
be located between the enhancer region and polyadenylation signal
sequence region. In one embodiment, a viral genome may comprise two
filler sequences, the first filler sequence may be located between
the payload region and exon region, and the second filler sequence
may be located between the enhancer region and MCS region. In one
embodiment, a viral genome may comprise two filler sequences, the
first filler sequence may be located between the payload region and
exon region, and the second filler sequence may be located between
the enhancer region and exon region. In one embodiment, a viral
genome may comprise two filler sequences, the first filler sequence
may be located between the payload region and exon region, and the
second filler sequence may be located between the enhancer region
and 3' ITR. In one embodiment, a viral genome may comprise two
filler sequences, the first filler sequence may be located between
the payload region and exon region, and the second filler sequence
may be located between the polyadenylation signal sequence region
and MCS region. In one embodiment, a viral genome may comprise two
filler sequences, the first filler sequence may be located between
the payload region and exon region, and the second filler sequence
may be located between the polyadenylation signal sequence region
and exon region. In one embodiment, a viral genome may comprise two
filler sequences, the first filler sequence may be located between
the payload region and exon region, and the second filler sequence
may be located between the polyadenylation signal sequence region
and 3' ITR. In one embodiment, a viral genome may comprise two
filler sequences, the first filler sequence may be located between
the payload region and exon region, and the second filler sequence
may be located between the MCS region and exon region. In one
embodiment, a viral genome may comprise two filler sequences, the
first filler sequence may be located between the payload region and
exon region, and the second filler sequence may be located between
the MCS region and 3' ITR. In one embodiment, a viral genome may
comprise two filler sequences, the first filler sequence may be
located between the payload region and exon region, and the second
filler sequence may be located between the exon region and 3'
ITR.
[0202] In one embodiment, a viral genome may comprise two filler
sequences, the first filler sequence may be located between the
payload region and 3' ITR region, and the second filler sequence
may be located between the intron region and enhancer region. In
one embodiment, a viral genome may comprise two filler sequences,
the first filler sequence may be located between the payload region
and 3' ITR region, and the second filler sequence may be located
between the intron region and polyadenylation signal sequence
region. In one embodiment, a viral genome may comprise two filler
sequences, the first filler sequence may be located between the
payload region and 3' ITR region, and the second filler sequence
may be located between the intron region and MCS region. In one
embodiment, a viral genome may comprise two filler sequences, the
first filler sequence may be located between the payload region and
3' ITR region, and the second filler sequence may be located
between the intron region and exon region. In one embodiment, a
viral genome may comprise two filler sequences, the first filler
sequence may be located between the payload region and 3' ITR
region, and the second filler sequence may be located between the
intron region and 3' ITR. In one embodiment, a viral genome may
comprise two filler sequences, the first filler sequence may be
located between the payload region and 3' ITR region, and the
second filler sequence may be located between the enhancer region
and polyadenylation signal sequence region. In one embodiment, a
viral genome may comprise two filler sequences, the first filler
sequence may be located between the payload region and 3' ITR
region, and the second filler sequence may be located between the
enhancer region and MCS region. In one embodiment, a viral genome
may comprise two filler sequences, the first filler sequence may be
located between the payload region and 3' ITR region, and the
second filler sequence may be located between the enhancer region
and exon region. In one embodiment, a viral genome may comprise two
filler sequences, the first filler sequence may be located between
the payload region and 3' ITR region, and the second filler
sequence may be located between the enhancer region and 3' ITR. In
one embodiment, a viral genome may comprise two filler sequences,
the first filler sequence may be located between the payload region
and 3' ITR region, and the second filler sequence may be located
between the polyadenylation signal sequence region and MCS region.
In one embodiment, a viral genome may comprise two filler
sequences, the first filler sequence may be located between the
payload region and 3' ITR region, and the second filler sequence
may be located between the polyadenylation signal sequence region
and exon region. In one embodiment, a viral genome may comprise two
filler sequences, the first filler sequence may be located between
the payload region and 3' ITR region, and the second filler
sequence may be located between the polyadenylation signal sequence
region and 3' ITR. In one embodiment, a viral genome may comprise
two filler sequences, the first filler sequence may be located
between the payload region and 3' ITR region, and the second filler
sequence may be located between the MCS region and exon region. In
one embodiment, a viral genome may comprise two filler sequences,
the first filler sequence may be located between the payload region
and 3' ITR region, and the second filler sequence may be located
between the MCS region and 3' ITR. In one embodiment, a viral
genome may comprise two filler sequences, the first filler sequence
may be located between the payload region and 3' ITR region, and
the second filler sequence may be located between the exon region
and 3' ITR.
[0203] In one embodiment, a viral genome may comprise two filler
sequences, the first filler sequence may be located between the
intron region and enhancer region, and the second filler sequence
may be located between the enhancer region and polyadenylation
signal sequence region. In one embodiment, a viral genome may
comprise two filler sequences, the first filler sequence may be
located between the intron region and enhancer region, and the
second filler sequence may be located between the enhancer region
and MCS region. In one embodiment, a viral genome may comprise two
filler sequences, the first filler sequence may be located between
the intron region and enhancer region, and the second filler
sequence may be located between the enhancer region and exon
region. In one embodiment, a viral genome may comprise two filler
sequences, the first filler sequence may be located between the
intron region and enhancer region, and the second filler sequence
may be located between the enhancer region and 3' ITR. In one
embodiment, a viral genome may comprise two filler sequences, the
first filler sequence may be located between the intron region and
enhancer region, and the second filler sequence may be located
between the polyadenylation signal sequence region and MCS region.
In one embodiment, a viral genome may comprise two filler
sequences, the first filler sequence may be located between the
intron region and enhancer region, and the second filler sequence
may be located between the polyadenylation signal sequence region
and exon region. In one embodiment, a viral genome may comprise two
filler sequences, the first filler sequence may be located between
the intron region and enhancer region, and the second filler
sequence may be located between the polyadenylation signal sequence
region and 3' ITR. In one embodiment, a viral genome may comprise
two filler sequences, the first filler sequence may be located
between the intron region and enhancer region, and the second
filler sequence may be located between the MCS region and exon
region. In one embodiment, a viral genome may comprise two filler
sequences, the first filler sequence may be located between the
intron region and enhancer region, and the second filler sequence
may be located between the MCS region and 3' ITR. In one
embodiment, a viral genome may comprise two filler sequences, the
first filler sequence may be located between the intron region and
enhancer region, and the second filler sequence may be located
between the exon region and 3' ITR.
[0204] In one embodiment, a viral genome may comprise two filler
sequences, the first filler sequence may be located between the
intron region and polyadenylation signal sequence region, and the
second filler sequence may be located between the enhancer region
and polyadenylation signal sequence region. In one embodiment, a
viral genome may comprise two filler sequences, the first filler
sequence may be located between the intron region and
polyadenylation signal sequence region, and the second filler
sequence may be located between the enhancer region and MCS region.
In one embodiment, a viral genome may comprise two filler
sequences, the first filler sequence may be located between the
intron region and polyadenylation signal sequence region, and the
second filler sequence may be located between the enhancer region
and exon region. In one embodiment, a viral genome may comprise two
filler sequences, the first filler sequence may be located between
the intron region and polyadenylation signal sequence region, and
the second filler sequence may be located between the enhancer
region and 3' ITR. In one embodiment, a viral genome may comprise
two filler sequences, the first filler sequence may be located
between the intron region and polyadenylation signal sequence
region, and the second filler sequence may be located between the
polyadenylation signal sequence region and MCS region. In one
embodiment, a viral genome may comprise two filler sequences, the
first filler sequence may be located between the intron region and
polyadenylation signal sequence region, and the second filler
sequence may be located between the polyadenylation signal sequence
region and exon region. In one embodiment, a viral genome may
comprise two filler sequences, the first filler sequence may be
located between the intron region and polyadenylation signal
sequence region, and the second filler sequence may be located
between the polyadenylation signal sequence region and 3' ITR. In
one embodiment, a viral genome may comprise two filler sequences,
the first filler sequence may be located between the intron region
and polyadenylation signal sequence region, and the second filler
sequence may be located between the MCS region and exon region. In
one embodiment, a viral genome may comprise two filler sequences,
the first filler sequence may be located between the intron region
and polyadenylation signal sequence region, and the second filler
sequence may be located between the MCS region and 3' ITR. In one
embodiment, a viral genome may comprise two filler sequences, the
first filler sequence may be located between the intron region and
polyadenylation signal sequence region, and the second filler
sequence may be located between the exon region and 3' ITR.
[0205] In one embodiment, a viral genome may comprise two filler
sequences, the first filler sequence may be located between the
intron region and MCS region, and the second filler sequence may be
located between the enhancer region and polyadenylation signal
sequence region. In one embodiment, a viral genome may comprise two
filler sequences, the first filler sequence may be located between
the intron region and MCS region, and the second filler sequence
may be located between the enhancer region and MCS region. In one
embodiment, a viral genome may comprise two filler sequences, the
first filler sequence may be located between the intron region and
MCS region, and the second filler sequence may be located between
the enhancer region and exon region. In one embodiment, a viral
genome may comprise two filler sequences, the first filler sequence
may be located between the intron region and MCS region, and the
second filler sequence may be located between the enhancer region
and 3' ITR. In one embodiment, a viral genome may comprise two
filler sequences, the first filler sequence may be located between
the intron region and MCS region, and the second filler sequence
may be located between the polyadenylation signal sequence region
and MCS region. In one embodiment, a viral genome may comprise two
filler sequences, the first filler sequence may be located between
the intron region and MCS region, and the second filler sequence
may be located between the polyadenylation signal sequence region
and exon region. In one embodiment, a viral genome may comprise two
filler sequences, the first filler sequence may be located between
the intron region and MCS region, and the second filler sequence
may be located between the polyadenylation signal sequence region
and 3' ITR. In one embodiment, a viral genome may comprise two
filler sequences, the first filler sequence may be located between
the intron region and MCS region, and the second filler sequence
may be located between the MCS region and exon region. In one
embodiment, a viral genome may comprise two filler sequences, the
first filler sequence may be located between the intron region and
MCS region, and the second filler sequence may be located between
the MCS region and 3' ITR. In one embodiment, a viral genome may
comprise two filler sequences, the first filler sequence may be
located between the intron region and MCS region, and the second
filler sequence may be located between the exon region and 3'
ITR.
[0206] In one embodiment, a viral genome may comprise two filler
sequences, the first filler sequence may be located between the
intron region and exon region, and the second filler sequence may
be located between the enhancer region and polyadenylation signal
sequence region. In one embodiment, a viral genome may comprise two
filler sequences, the first filler sequence may be located between
the intron region and exon region, and the second filler sequence
may be located between the enhancer region and MCS region. In one
embodiment, a viral genome may comprise two filler sequences, the
first filler sequence may be located between the intron region and
exon region, and the second filler sequence may be located between
the enhancer region and exon region. In one embodiment, a viral
genome may comprise two filler sequences, the first filler sequence
may be located between the intron region and exon region, and the
second filler sequence may be located between the enhancer region
and 3' ITR. In one embodiment, a viral genome may comprise two
filler sequences, the first filler sequence may be located between
the intron region and exon region, and the second filler sequence
may be located between the polyadenylation signal sequence region
and MCS region. In one embodiment, a viral genome may comprise two
filler sequences, the first filler sequence may be located between
the intron region and exon region, and the second filler sequence
may be located between the polyadenylation signal sequence region
and exon region. In one embodiment, a viral genome may comprise two
filler sequences, the first filler sequence may be located between
the intron region and exon region, and the second filler sequence
may be located between the polyadenylation signal sequence region
and 3' ITR. In one embodiment, a viral genome may comprise two
filler sequences, the first filler sequence may be located between
the intron region and exon region, and the second filler sequence
may be located between the MCS region and exon region. In one
embodiment, a viral genome may comprise two filler sequences, the
first filler sequence may be located between the intron region and
exon region, and the second filler sequence may be located between
the MCS region and 3' ITR. In one embodiment, a viral genome may
comprise two filler sequences, the first filler sequence may be
located between the intron region and exon region, and the second
filler sequence may be located between the exon region and 3'
ITR.
[0207] In one embodiment, a viral genome may comprise two filler
sequences, the first filler sequence may be located between the
intron region and 3'ITR, and the second filler sequence may be
located between the enhancer region and polyadenylation signal
sequence region. In one embodiment, a viral genome may comprise two
filler sequences, the first filler sequence may be located between
the intron region and 3'ITR, and the second filler sequence may be
located between the enhancer region and MCS region. In one
embodiment, a viral genome may comprise two filler sequences, the
first filler sequence may be located between the intron region and
3'ITR, and the second filler sequence may be located between the
enhancer region and exon region. In one embodiment, a viral genome
may comprise two filler sequences, the first filler sequence may be
located between the intron region and 3'ITR, and the second filler
sequence may be located between the enhancer region and 3' ITR. In
one embodiment, a viral genome may comprise two filler sequences,
the first filler sequence may be located between the intron region
and 3'ITR, and the second filler sequence may be located between
the polyadenylation signal sequence region and MCS region. In one
embodiment, a viral genome may comprise two filler sequences, the
first filler sequence may be located between the intron region and
3'ITR, and the second filler sequence may be located between the
polyadenylation signal sequence region and exon region. In one
embodiment, a viral genome may comprise two filler sequences, the
first filler sequence may be located between the intron region and
3'ITR, and the second filler sequence may be located between the
polyadenylation signal sequence region and 3' ITR. In one
embodiment, a viral genome may comprise two filler sequences, the
first filler sequence may be located between the intron region and
3'ITR, and the second filler sequence may be located between the
MCS region and exon region. In one embodiment, a viral genome may
comprise two filler sequences, the first filler sequence may be
located between the intron region and 3'ITR, and the second filler
sequence may be located between the MCS region and 3' ITR. In one
embodiment, a viral genome may comprise two filler sequences, the
first filler sequence may be located between the intron region and
3'ITR, and the second filler sequence may be located between the
exon region and 3' ITR.
[0208] In one embodiment, a viral genome may comprise two filler
sequences, the first filler sequence may be located between the
enhancer region and polyadenylation signal sequence region, and the
second filler sequence may be located between the polyadenylation
signal sequence region and MCS region. In one embodiment, a viral
genome may comprise two filler sequences, the first filler sequence
may be located between the enhancer region and polyadenylation
signal sequence region, and the second filler sequence may be
located between the polyadenylation signal sequence region and exon
region. In one embodiment, a viral genome may comprise two filler
sequences, the first filler sequence may be located between the
enhancer region and polyadenylation signal sequence region, and the
second filler sequence may be located between the polyadenylation
signal sequence region and 3' ITR. In one embodiment, a viral
genome may comprise two filler sequences, the first filler sequence
may be located between the enhancer region and polyadenylation
signal sequence region, and the second filler sequence may be
located between the MCS region and exon region. In one embodiment,
a viral genome may comprise two filler sequences, the first filler
sequence may be located between the enhancer region and
polyadenylation signal sequence region, and the second filler
sequence may be located between the MCS region and 3' ITR. In one
embodiment, a viral genome may comprise two filler sequences, the
first filler sequence may be located between the enhancer region
and polyadenylation signal sequence region, and the second filler
sequence may be located between the exon region and 3' ITR.
[0209] In one embodiment, a viral genome may comprise two filler
sequences, the first filler sequence may be located between the
enhancer region and MCS region, and the second filler sequence may
be located between the polyadenylation signal sequence region and
MCS region. In one embodiment, a viral genome may comprise two
filler sequences, the first filler sequence may be located between
the enhancer region and MCS region, and the second filler sequence
may be located between the polyadenylation signal sequence region
and exon region. In one embodiment, a viral genome may comprise two
filler sequences, the first filler sequence may be located between
the enhancer region and MCS region, and the second filler sequence
may be located between the polyadenylation signal sequence region
and 3' ITR. In one embodiment, a viral genome may comprise two
filler sequences, the first filler sequence may be located between
the enhancer region and MCS region, and the second filler sequence
may be located between the MCS region and exon region. In one
embodiment, a viral genome may comprise two filler sequences, the
first filler sequence may be located between the enhancer region
and MCS region, and the second filler sequence may be located
between the MCS region and 3' ITR. In one embodiment, a viral
genome may comprise two filler sequences, the first filler sequence
may be located between the enhancer region and MCS region, and the
second filler sequence may be located between the exon region and
3' ITR.
[0210] In one embodiment, a viral genome may comprise two filler
sequences, the first filler sequence may be located between the
enhancer region and exon region, and the second filler sequence may
be located between the polyadenylation signal sequence region and
MCS region. In one embodiment, a viral genome may comprise two
filler sequences, the first filler sequence may be located between
the enhancer region and exon region, and the second filler sequence
may be located between the polyadenylation signal sequence region
and exon region. In one embodiment, a viral genome may comprise two
filler sequences, the first filler sequence may be located between
the enhancer region and exon region, and the second filler sequence
may be located between the polyadenylation signal sequence region
and 3' ITR. In one embodiment, a viral genome may comprise two
filler sequences, the first filler sequence may be located between
the enhancer region and exon region, and the second filler sequence
may be located between the MCS region and exon region. In one
embodiment, a viral genome may comprise two filler sequences, the
first filler sequence may be located between the enhancer region
and exon region, and the second filler sequence may be located
between the MCS region and 3' ITR. In one embodiment, a viral
genome may comprise two filler sequences, the first filler sequence
may be located between the enhancer region and exon region, and the
second filler sequence may be located between the exon region and
3' ITR.
[0211] In one embodiment, a viral genome may comprise two filler
sequences, the first filler sequence may be located between the
enhancer region and 3' ITR, and the second filler sequence may be
located between the polyadenylation signal sequence region and MCS
region. In one embodiment, a viral genome may comprise two filler
sequences, the first filler sequence may be located between the
enhancer region and 3' ITR, and the second filler sequence may be
located between the polyadenylation signal sequence region and exon
region. In one embodiment, a viral genome may comprise two filler
sequences, the first filler sequence may be located between the
enhancer region and 3' ITR, and the second filler sequence may be
located between the polyadenylation signal sequence region and 3'
ITR. In one embodiment, a viral genome may comprise two filler
sequences, the first filler sequence may be located between the
enhancer region and 3' ITR, and the second filler sequence may be
located between the MCS region and exon region. In one embodiment,
a viral genome may comprise two filler sequences, the first filler
sequence may be located between the enhancer region and 3' ITR, and
the second filler sequence may be located between the MCS region
and 3' ITR. In one embodiment, a viral genome may comprise two
filler sequences, the first filler sequence may be located between
the enhancer region and 3' ITR, and the second filler sequence may
be located between the exon region and 3' ITR.
[0212] In one embodiment, a viral genome may comprise two filler
sequences, the first filler sequence may be located between the
polyadenylation signal sequence region and MCS region, and the
second filler sequence may be located between the MCS region and
exon region. In one embodiment, a viral genome may comprise two
filler sequences, the first filler sequence may be located between
the polyadenylation signal sequence region and MCS region, and the
second filler sequence may be located between the MCS region and 3'
ITR. In one embodiment, a viral genome may comprise two filler
sequences, the first filler sequence may be located between the
polyadenylation signal sequence region and MCS region, and the
second filler sequence may be located between the exon region and
3' ITR.
[0213] In one embodiment, a viral genome may comprise two filler
sequences, the first filler sequence may be located between the
polyadenylation signal sequence region and exon region, and the
second filler sequence may be located between the MCS region and
exon region. In one embodiment, a viral genome may comprise two
filler sequences, the first filler sequence may be located between
the polyadenylation signal sequence region and exon region, and the
second filler sequence may be located between the MCS region and 3'
ITR. In one embodiment, a viral genome may comprise two filler
sequences, the first filler sequence may be located between the
polyadenylation signal sequence region and exon region, and the
second filler sequence may be located between the exon region and
3' ITR.
[0214] In one embodiment, a viral genome may comprise two filler
sequences, the first filler sequence may be located between the
polyadenylation signal sequence region and 3' ITR, and the second
filler sequence may be located between the MCS region and exon
region. In one embodiment, a viral genome may comprise two filler
sequences, the first filler sequence may be located between the
polyadenylation signal sequence region and 3' ITR, and the second
filler sequence may be located between the MCS region and 3' ITR.
In one embodiment, a viral genome may comprise two filler
sequences, the first filler sequence may be located between the
polyadenylation signal sequence region and 3' ITR, and the second
filler sequence may be located between the exon region and 3'
ITR.
[0215] In one embodiment, a viral genome may comprise two filler
sequences, the first filler sequence may be located between the MCS
region and exon region, and the second filler sequence may be
located between the exon region and 3' ITR.
AAV Production
[0216] The present disclosure provides methods for the generation
of parvoviral particles, e.g. AAV particles, by viral genome
replication in a viral replication cell.
[0217] In accordance with the disclosure, the viral genome
comprising a payload region will be incorporated into the AAV
particle produced in the viral replication cell. Methods of making
AAV particles are well known in the art and are described in e.g.,
U.S. Pat. No. 6,204,059, U.S. Pat. No. 5,756,283, U.S. Pat. No.
6,258,595, U.S. Pat. No. 6,261,551, U.S. Pat. No. 6,270,996, U.S.
Pat. No. 6,281,010, U.S. Pat. No. 6,365,394, U.S. Pat. No.
6,475,769, U.S. Pat. No. 6,482,634, U.S. Pat. No. 6,485,966, U.S.
Pat. No. 6,943,019, U.S. Pat. No. 6,953,690, U.S. Pat. No.
7,022,519, U.S. Pat. No. 7,238,526, U.S. Pat. No. 7,291,498 and
U.S. Pat. No. 7,491,508, U.S. Pat. No. 5,064,764, U.S. Pat. No.
6,194,191, U.S. Pat. No. 6,566,118, U.S. Pat. No. 8,137,948; or
International Publication Nos. WO1996039530, WO1998010088,
WO1999014354, WO1999015685, WO1999047691, WO2000055342,
WO2000075353 and WO2001023597; Methods In Molecular Biology, ed.
Richard, Humana Press, N J (1995); O'Reilly et al., Baculovirus
Expression Vectors, A Laboratory Manual, Oxford Univ. Press (1994);
Samulski et al., J. Vir. 63:3822-8 (1989); Kajigaya et al., Proc.
Nat'l. Acad. Sci. USA 88: 4646-50 (1991); Ruffing et al., J. Vir.
66:6922-30 (1992); Kimbauer et al., Vir., 219:37-44 (1996); Zhao et
al., Vir. 272:382-93 (2000); the contents of each of which are
herein incorporated by reference in their entirety. In one
embodiment, the AAV particles are made using the methods described
in WO2015191508, the contents of which are herein incorporated by
reference in their entirety.
[0218] Viral replication cells commonly used for production of
recombinant AAV particles include but are not limited to 293 cells,
COS cells, HeLa cells, KB cells, and other mammalian cell lines as
described in U.S. Pat. No. 6,156,303, U.S. Pat. No. 5,387,484, U.S.
Pat. No. 5,741,683, U.S. Pat. No. 5,691,176, and U.S. Pat. No.
5,688,676; U.S. patent publication No. 2002/0081721, and
International Patent Publication Nos. WO 00/47757, WO 00/24916, and
WO 96/17947, the contents of each of which are herein incorporated
by reference in their entireties.
[0219] In some embodiments, the present disclosure provides a
method for producing an AAV particle having enhanced (increased,
improved) transduction efficiency comprising the steps of: 1)
co-transfecting competent bacterial cells with a bacmid vector and
either a viral construct vector and/or AAV payload construct
vector, 2) isolating the resultant viral construct expression
vector and AAV payload construct expression vector and separately
transfecting viral replication cells, 3) isolating and purifying
resultant payload and viral construct particles comprising viral
construct expression vector or AAV payload construct expression
vector, 4) co-infecting a viral replication cell with both the AAV
payload and viral construct particles comprising viral construct
expression vector or AAV payload construct expression vector, and
5) harvesting and purifying the AAV particle comprising a viral
genome.
[0220] In some embodiments, the present disclosure provides a
method for producing an AAV particle comprising the steps of 1)
simultaneously co-transfecting mammalian cells, such as, but not
limited to HEK293 cells, with a payload region, a construct
expressing rep and cap genes and a helper construct, 2) harvesting
and purifying the AAV particle comprising a viral genome.
[0221] In some embodiments, the viral genome of the AAV particle of
the disclosure optionally encodes a selectable marker. The
selectable marker may comprise a cell-surface marker, such as any
protein expressed on the surface of the cell including, but not
limited to receptors, CD markers, lectins, integrins, or truncated
versions thereof.
[0222] In some embodiments, selectable marker reporter genes as
described in International application No. WO 96/23810; Heim et
al., Current Biology 2:178-182 (1996); Heim et al., Proc. Natl.
Acad. Sci. USA (1995); or Heim et al., Science 373:663-664 (1995);
WO 96/30540, the contents of each of which are incorporated herein
by reference in their entireties).
Genome Size
[0223] In one embodiment, the AAV particle which comprises a
payload described herein may be single stranded or double stranded
vector genome. The size of the vector genome may be small, medium,
large or the maximum size. Additionally, the vector genome may
comprise a promoter and a polyA tail.
[0224] In one embodiment, the vector genome which comprises a
payload described herein may be a small single stranded vector
genome. A small single stranded vector genome may be 2.7 to 3.5 kb
in size such as about 2.7, 2.8, 2.9, 3.0, 3.1, 3.2, 3.3, 3.4, and
3.5 kb in size. As a non-limiting example, the small single
stranded vector genome may be 3.2 kb in size. Additionally, the
vector genome may comprise a promoter and a polyA tail.
[0225] In one embodiment, the vector genome which comprises a
payload described herein may be a small double stranded vector
genome. A small double stranded vector genome may be 1.3 to 1.7 kb
in size such as about 1.3, 1.4, 1.5, 1.6, and 1.7 kb in size. As a
non-limiting example, the small double stranded vector genome may
be 1.6 kb in size. Additionally, the vector genome may comprise a
promoter and a polyA tail.
[0226] In one embodiment, the vector genome which comprises a
payload described herein may be a medium single stranded vector
genome. A medium single stranded vector genome may be 3.6 to 4.3 kb
in size such as about 3.6, 3.7, 3.8, 3.9, 4.0, 4.1, 4.2 and 4.3 kb
in size. As a non-limiting example, the medium single stranded
vector genome may be 4.0 kb in size. Additionally, the vector
genome may comprise a promoter and a polyA tail.
[0227] In one embodiment, the vector genome which comprises a
payload described herein may be a medium double stranded vector
genome. A medium double stranded vector genome may be 1.8 to 2.1 kb
in size such as about 1.8, 1.9, 2.0, and 2.1 kb in size. As a
non-limiting example, the medium double stranded vector genome may
be 2.0 kb in size. Additionally, the vector genome may comprise a
promoter and a polyA tail.
[0228] In one embodiment, the vector genome which comprises a
payload described herein may be a large single stranded vector
genome. A large single stranded vector genome may be 4.4 to 6.0 kb
in size such as about 4.4, 4.5, 4.6, 4.7, 4.8, 4.9, 5.0, 5.1, 5.2,
5.3, 5.4, 5.5, 5.6, 5.7, 5.8, 5.9 and 6.0 kb in size. As a
non-limiting example, the large single stranded vector genome may
be 4.7 kb in size. As another non-limiting example, the large
single stranded vector genome may be 4.8 kb in size. As yet another
non-limiting example, the large single stranded vector genome may
be 6.0 kb in size. Additionally, the vector genome may comprise a
promoter and a polyA tail.
[0229] In one embodiment, the vector genome which comprises a
payload described herein may be a large double stranded vector
genome. A large double stranded vector genome may be 2.2 to 3.0 kb
in size such as about 2.2, 2.3, 2.4, 2.5, 2.6, 2.7, 2.8, 2.9 and
3.0 kb in size. As a non-limiting example, the large double
stranded vector genome may be 2.4 kb in size. Additionally, the
vector genome may comprise a promoter and a polyA tail.
Payloads
[0230] The AAV particles of the present disclosure comprise at
least one payload region. As used herein, "payload" or "payload
region" refers to one or more polynucleotides or polynucleotide
regions encoded by or within a viral genome or an expression
product of such polynucleotide or polynucleotide region, e.g., a
transgene, a polynucleotide encoding a polypeptide or
multi-polypeptide or a modulatory nucleic acid or regulatory
nucleic acid. Payloads of the present disclosure typically encode
polypeptides or fragments or variants thereof.
[0231] The payload region may be constructed in such a way as to
reflect a region similar to or mirroring the natural organization
of an mRNA.
[0232] The payload region may comprise a combination of coding and
non-coding nucleic acid sequences.
[0233] In some embodiments, the AAV payload region may encode a
coding or non-coding RNA.
[0234] In one embodiment, the AAV particle comprises a viral genome
with a payload region comprising nucleic acid sequences encoding
more than one polypeptide of interest. In such an embodiment, a
viral genome encoding more than one polypeptide may be replicated
and packaged into a viral particle. A target cell transduced with a
viral particle comprising more than one polypeptide may express
each of the polypeptides in a single cell.
[0235] In one embodiment, the payload region may comprise the
components as shown in FIG. 1. The payload region 110 is located
within the viral genome 100. At the 5' and/or the 3' end of the
payload region 110 there may be at least one inverted terminal
repeat (ITR) 120. Within the payload region, there is a promoter
region 130, an intron region 140 and a coding region 150.
[0236] Where the AAV particle payload region encodes a polypeptide,
the polypeptide may be a peptide or protein. The viral genomes
encoding polypeptides described herein may be useful in the fields
of human disease, viruses, infections veterinary applications and a
variety of in vivo and in vitro settings.
[0237] In some embodiments, the AAV particles are useful in the
field of medicine for the treatment, prophylaxis, palliation or
amelioration of neurological diseases and/or disorders.
[0238] In some embodiments, the AAV particles are useful in the
field of medicine for the treatment, prophylaxis, palliation or
amelioration of Parkinson's Disease.
[0239] In some embodiments, the AAV particles are useful in the
field of medicine for the treatment, prophylaxis, palliation or
amelioration of diseases of the central nervous system.
The Nature of the Polypeptides and Variants
[0240] Amino acid sequences encoded by payload regions of the viral
genomes of the disclosure may be translated as a whole polypeptide,
a plurality of polypeptides or fragments of polypeptides, which
independently may be encoded by one or more nucleic acids,
fragments of nucleic acids or variants of any of the
aforementioned. As used herein, "polypeptide" means a polymer of
amino acid residues (natural or unnatural) linked together most
often by peptide bonds. The term, as used herein, refers to
proteins, polypeptides, and peptides of any size, structure, or
function. In some instances, the polypeptide encoded is smaller
than about 50 amino acids and the polypeptide is then termed a
peptide. If the polypeptide is a peptide, it will be at least about
2, 3, 4, or at least 5 amino acid residues long. Thus, polypeptides
include gene products, naturally occurring polypeptides, synthetic
polypeptides, homologs, orthologs, paralogs, fragments and other
equivalents, variants, and analogs of the foregoing. A polypeptide
may be a single molecule or may be a multi-molecular complex such
as a dimer, trimer or tetramer. They may also comprise single chain
or multichain polypeptides and may be associated or linked. The
term polypeptide may also apply to amino acid polymers in which one
or more amino acid residues are an artificial chemical analogue of
a corresponding naturally occurring amino acid.
[0241] The term "polypeptide variant" refers to molecules which
differ in their amino acid sequence from a native or reference
sequence. The amino acid sequence variants may possess
substitutions, deletions, and/or insertions at certain positions
within the amino acid sequence, as compared to a native or
reference sequence. Ordinarily, variants will possess at least
about 50% identity (homology) to a native or reference sequence,
and preferably, they will be at least about 80%, more preferably at
least about 90% identical (homologous) to a native or reference
sequence.
[0242] In some embodiments "variant mimics" are provided. As used
herein, the term "variant mimic" is one which contains one or more
amino acids which would mimic an activated sequence. For example,
glutamate may serve as a mimic for phosphoro-threonine and/or
phosphoro-serine. Alternatively, variant mimics may result in
deactivation or in an inactivated product containing the mimic,
e.g., phenylalanine may act as an inactivating substitution for
tyrosine; or alanine may act as an inactivating substitution for
serine.
[0243] The term "amino acid sequence variant" refers to molecules
with some differences in their amino acid sequences as compared to
a native or starting sequence. The amino acid sequence variants may
possess substitutions, deletions, and/or insertions at certain
positions within the amino acid sequence. "Native" or "starting"
sequence should not be confused with a wild type sequence. As used
herein, a native or starting sequence is a relative term referring
to an original molecule against which a comparison may be made.
"Native" or "starting" sequences or molecules may represent the
wild-type (that sequence found in nature) but do not have to be the
wild-type sequence.
[0244] Ordinarily, variants will possess at least about 70%
homology to a native sequence, and preferably, they will be at
least about 80%, more preferably at least about 90% homologous to a
native sequence. "Homology" as it applies to amino acid sequences
is defined as the percentage of residues in the candidate amino
acid sequence that are identical with the residues in the amino
acid sequence of a second sequence after aligning the sequences and
introducing gaps, if necessary, to achieve the maximum percent
homology. Methods and computer programs for the alignment are well
known in the art. It is understood that homology depends on a
calculation of percent identity but may differ in value due to gaps
and penalties introduced in the calculation.
[0245] By "homologs" as it applies to amino acid sequences is meant
the corresponding sequence of other species having substantial
identity to a second sequence of a second species.
[0246] "Analogs" is meant to include polypeptide variants which
differ by one or more amino acid alterations, e.g., substitutions,
additions or deletions of amino acid residues that still maintain
the properties of the parent polypeptide.
[0247] Sequence tags or amino acids, such as one or more lysines,
can be added to the peptide sequences of the disclosure (e.g., at
the N-terminal or C-terminal ends). Sequence tags can be used for
peptide purification or localization. Lysines can be used to
increase peptide solubility or to allow for biotinylation.
Alternatively, amino acid residues located at the carboxy and amino
terminal regions of the amino acid sequence of a peptide or protein
may optionally be deleted providing for truncated sequences.
Certain amino acids (e.g., C-terminal or N-terminal residues) may
alternatively be deleted depending on the use of the sequence, as
for example, expression of the sequence as part of a larger
sequence which is soluble, or linked to a solid support.
[0248] "Substitutional variants" when referring to proteins are
those that have at least one amino acid residue in a native or
starting sequence removed and a different amino acid inserted in
its place at the same position. The substitutions may be single,
where only one amino acid in the molecule has been substituted, or
they may be multiple, where two or more amino acids have been
substituted in the same molecule.
[0249] As used herein the term "conservative amino acid
substitution" refers to the substitution of an amino acid that is
normally present in the sequence with a different amino acid of
similar size, charge, or polarity. Examples of conservative
substitutions include the substitution of a non-polar (hydrophobic)
residue such as isoleucine, valine and leucine for another
non-polar residue. Likewise, examples of conservative substitutions
include the substitution of one polar (hydrophilic) residue for
another such as between arginine and lysine, between glutamine and
asparagine, and between glycine and serine. Additionally, the
substitution of a basic residue such as lysine, arginine or
histidine for another, or the substitution of one acidic residue
such as aspartic acid or glutamic acid for another acidic residue
are additional examples of conservative substitutions. Examples of
non-conservative substitutions include the substitution of a
non-polar (hydrophobic) amino acid residue such as isoleucine,
valine, leucine, alanine, methionine for a polar (hydrophilic)
residue such as cysteine, glutamine, glutamic acid or lysine and/or
a polar residue for a non-polar residue.
[0250] "Insertional variants" when referring to proteins are those
with one or more amino acids inserted immediately adjacent to an
amino acid at a particular position in a native or starting
sequence. "Immediately adjacent" to an amino acid means connected
to either the alpha-carboxy or alpha-amino functional group of the
amino acid.
[0251] "Deletional variants" when referring to proteins, are those
with one or more amino acids in the native or starting amino acid
sequence removed. Ordinarily, deletional variants will have one or
more amino acids deleted in a particular region of the
molecule.
[0252] As used herein, the term "derivative" is used synonymously
with the term "variant" and refers to a molecule that has been
modified or changed in any way relative to a reference molecule or
starting molecule. In some embodiments, derivatives include native
or starting proteins that have been modified with an organic
proteinaceous or non-proteinaceous derivatizing agent, and
post-translational modifications. Covalent modifications are
traditionally introduced by reacting targeted amino acid residues
of the protein with an organic derivatizing agent that is capable
of reacting with selected side-chains or terminal residues, or by
harnessing mechanisms of post-translational modifications that
function in selected recombinant host cells. The resultant covalent
derivatives are useful in programs directed at identifying residues
important for biological activity, for immunoassays, or for the
preparation of anti-protein antibodies for immunoaffinity
purification of the recombinant glycoprotein. Such modifications
are within the ordinary skill in the art and are performed without
undue experimentation.
[0253] Certain post-translational modifications are the result of
the action of recombinant host cells on the expressed polypeptide.
Glutaminyl and asparaginyl residues are frequently
post-translationally deamidated to the corresponding glutamyl and
aspartyl residues. Alternatively, these residues are deamidated
under mildly acidic conditions. Either form of these residues may
be present in the proteins used in accordance with the present
disclosure.
[0254] Other post-translational modifications include hydroxylation
of proline and lysine, phosphorylation of hydroxyl groups of seryl
or threonyl residues, methylation of the alpha-amino groups of
lysine, arginine, and histidine side chains (T. E. Creighton,
Proteins: Structure and Molecular Properties, W.H. Freeman &
Co., San Francisco, pp. 79-86 (1983)).
[0255] "Features" when referring to proteins are defined as
distinct amino acid sequence-based components of a molecule.
Features of the proteins of the present disclosure include surface
manifestations, local conformational shape, folds, loops,
half-loops, domains, half-domains, sites, termini or any
combination thereof.
[0256] As used herein when referring to proteins the term "surface
manifestation" refers to a polypeptide based component of a protein
appearing on an outermost surface.
[0257] As used herein when referring to proteins the term "local
conformational shape" means a polypeptide based structural
manifestation of a protein which is located within a definable
space of the protein.
[0258] As used herein when referring to proteins the term "fold"
means the resultant conformation of an amino acid sequence upon
energy minimization. A fold may occur at the secondary or tertiary
level of the folding process. Examples of secondary level folds
include beta sheets and alpha helices. Examples of tertiary folds
include domains and regions formed due to aggregation or separation
of energetic forces. Regions formed in this way include hydrophobic
and hydrophilic pockets, and the like.
[0259] As used herein the term "turn" as it relates to protein
conformation means a bend which alters the direction of the
backbone of a peptide or polypeptide and may involve one, two,
three or more amino acid residues.
[0260] As used herein when referring to proteins the term "loop"
refers to a structural feature of a peptide or polypeptide which
reverses the direction of the backbone of a peptide or polypeptide
and comprises four or more amino acid residues. Oliva et al. have
identified at least 5 classes of protein loops (J. Mol Biol 266
(4): 814-830; 1997).
[0261] As used herein when referring to proteins the term
"half-loop" refers to a portion of an identified loop having at
least half the number of amino acid residues as the loop from which
it is derived. It is understood that loops may not always contain
an even number of amino acid residues. Therefore, in those cases
where a loop contains or is identified to comprise an odd number of
amino acids, a half-loop of the odd-numbered loop will comprise the
whole number portion or next whole number portion of the loop
(number of amino acids of the loop/2+/-0.5 amino acids). For
example, a loop identified as a 7 amino acid loop could produce
half-loops of 3 amino acids or 4 amino acids (7/2=3.5+/-0.5 being 3
or 4).
[0262] As used herein when referring to proteins the term "domain"
refers to a motif of a polypeptide having one or more identifiable
structural or functional characteristics or properties (e.g.,
binding capacity, serving as a site for protein-protein
interactions).
[0263] As used herein when referring to proteins the term
"half-domain" means portion of an identified domain having at least
half the number of amino acid residues as the domain from which it
is derived. It is understood that domains may not always contain an
even number of amino acid residues. Therefore, in those cases where
a domain contains or is identified to comprise an odd number of
amino acids, a half-domain of the odd-numbered domain will comprise
the whole number portion or next whole number portion of the domain
(number of amino acids of the domain/2+/-0.5 amino acids). For
example, a domain identified as a 7 amino acid domain could produce
half-domains of 3 amino acids or 4 amino acids (7/2=3.5+/-0.5 being
3 or 4). It is also understood that sub-domains may be identified
within domains or half-domains, these subdomains possessing less
than all of the structural or functional properties identified in
the domains or half domains from which they were derived. It is
also understood that the amino acids that comprise any of the
domain types herein need not be contiguous along the backbone of
the polypeptide (i.e., nonadjacent amino acids may fold
structurally to produce a domain, half-domain or subdomain).
[0264] As used herein when referring to proteins the terms "site"
as it pertains to amino acid based embodiments is used synonymous
with "amino acid residue" and "amino acid side chain". A site
represents a position within a peptide or polypeptide that may be
modified, manipulated, altered, derivatized or varied within the
polypeptide based molecules of the present disclosure.
[0265] As used herein the terms "termini or terminus" when
referring to proteins refers to an extremity of a peptide or
polypeptide. Such extremity is not limited only to the first or
final site of the peptide or polypeptide but may include additional
amino acids in the terminal regions. The polypeptide based
molecules of the present disclosure may be characterized as having
both an N-terminus (terminated by an amino acid with a free amino
group (NH2)) and a C-terminus (terminated by an amino acid with a
free carboxyl group (COOH)). Proteins of the disclosure are in some
cases made up of multiple polypeptide chains brought together by
disulfide bonds or by non-covalent forces (multimers, oligomers).
These sorts of proteins will have multiple N- and C-termini.
Alternatively, the termini of the polypeptides may be modified such
that they begin or end, as the case may be, with a non-polypeptide
based moiety such as an organic conjugate.
[0266] Once any of the features have been identified or defined as
a component of a molecule of the disclosure, any of several
manipulations and/or modifications of these features may be
performed by moving, swapping, inverting, deleting, randomizing or
duplicating. Furthermore, it is understood that manipulation of
features may result in the same outcome as a modification to the
molecules of the disclosure. For example, a manipulation which
involves deleting a domain would result in the alteration of the
length of a molecule just as modification of a nucleic acid to
encode less than a full length molecule would.
[0267] Modifications and manipulations can be accomplished by
methods known in the art such as site directed mutagenesis. The
resulting modified molecules may then be tested for activity using
in vitro or in vivo assays such as those described herein or any
other suitable screening assay known in the art.
Payload: AADC Polynucleotide Constructs
[0268] According to the present disclosure, aromatic L-amino acid
decarboxylase (AADC; also known as dopa decarboxylase and DDC)
polynucleotides are provided which function alone or in combination
with additional nucleic acid sequence(s) to encode the AADC
protein. As used herein an "AADC polynucleotide" is any nucleic
acid polymer which encodes an AADC protein and when present in a
vector, plasmid or translatable construct, expresses such AADC
protein in a cell, tissue, organ or organism.
[0269] AADC polynucleotides include precursor molecules which are
processed inside the cell. AADC polynucleotides or the processed
forms thereof may be encoded in a plasmid, vector, genome or other
nucleic acid expression vector for delivery to a cell.
[0270] In some embodiments AADC polynucleotides are designed as
components of AAV viral genomes and packaged in AAV particles which
are processed within the cell to produce the wild type AADC
protein.
[0271] In some embodiments, the AADC polynucleotide may be the
payload of the AAV particle.
[0272] As used herein, the wild type AADC protein may be any of the
naturally occurring isoforms or variants from the DDC gene.
Multiple alternatively spliced transcript variants encoding
different isoforms of AADC have been identified. Specifically, the
DDC gene produces seven transcript variants that encode six
distinct isoforms. DDC transcript variants 1 and 2 both encode AADC
isoform 1. In some embodiments, the AADC polynucleotides encode DDC
transcript variant 2, thereby encoding a native AADC isoform 1
(NCBI Reference Sequence: NP_000781.1). This sequence is given
here:
TABLE-US-00002 (SEQ ID NO: 978)
MNASEFRRRGKEMVDYVANYMEGIEGRQVYPDVEPGYLRPLIPAAAPQEP
DTFEDIINDVEKIIMPGVTHWHSPYFFAYFPTASSYPAMLADMLCGAIGC
IGFSWAASPACTELETVMMDWLGKMLELPKAFLNEKAGEGGGVIQGSASE
ATLVALLAARTKVIHRLQAASPELTQAAIMEKLVAYSSDQAHSSVERAGL
IGGVKLKAIPSDGNFAMRASALQEALERDKAAGLIPFFMVATLGTTTCCS
FDNLLEVGPICNKEDIWLHVDAAYAGSAFICPEFRHLLNGVEFADSFNFN
PHKWLLVNFDCSAMWVKKRTDLTGAFRLDPTYLKHSHQDSGLITDYRHWQ
IPLGRRFRSLKMWFVFRMYGVKGLQAYIRKHVQLSHEFESLVRQDPRFEI
CVEVILGLVCFRLKGSNKVNEALLQRINSAKKIHLVPCHLRDKFVLRFAI
CSRTVESAHVQRAWEHIKELAADVLRAERE
[0273] The AADC polynucleotides of the disclosure, may be
engineered to contain modular elements and/or sequence motifs
assembled to create AADC polynucleotide constructs.
[0274] According to the present disclosure, AADC polynucleotides
are provided. Such polynucleotides comprise nucleic acid polymers
which comprise a region of linked nucleosides encoding one or more
isoforms or variants of the AADC protein.
[0275] In some embodiments, the AADC polynucleotide comprises a
codon optimized transcript encoding an AADC protein.
[0276] In some embodiments, the AADC polynucleotide comprises a
sequence region encoding one or more wild type isoforms or variants
of an AADC protein. Such polynucleotides may also comprise a
sequence region encoding any one or more of the following: a 5'
ITR, a cytomegalovirus (CMV) Enhancer, a CMV Promoter, an ie1 exon
1, an ie1 intron1, an hbBglobin intron2, an hBglobin exon 3, a 5'
UTR, a 3' UTR, an hGH poly(A) signal, and/or a 3' ITR. Such
sequence regions are taught herein or may be any of those known in
the art.
[0277] In some embodiments, the AADC polynucleotide comprises a SEQ
ID NO: 979 or a fragment or variant thereof.
[0278] In one embodiment, the AADC polynucleotide comprises a
sequence which has a percent identity to any of SEQ ID NO: 979 or a
fragment or variant thereof. The AADC polynucleotide may have 1%,
2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 15%, 20%, 25%, 30%, 35%, 40%,
45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 99% or 100%
identity to any of SEQ ID NO: 979 or a fragment or variant thereof.
The AADC polynucleotide may have 1-10%, 10-20%, 30-40%, 50-60%,
50-70%, 50-80%, 50-90%, 50-99%, 50-100%, 60-70%, 60-80%, 60-90%,
60-99%, 60-100%, 70-80%, 70-90%, 70-99%, 70-100%, 80-85%, 80-90%,
80-95%, 80-99%, 80-100%, 90-95%, 90-99%, or 90-100% to any of SEQ
ID NO: 979 or a fragment or variant thereof. As a non-limiting
example, the AADC polynucleotide comprises a sequence which as 80%
identity to any of SEQ ID NO: 979 or a fragment or variant thereof.
As another non-limiting example, the AADC polynucleotide comprises
a sequence which as 85% identity to any of SEQ ID NO: 979 or a
fragment or variant thereof. As another non-limiting example, the
AADC polynucleotide comprises a sequence which as 90% identity to
any of SEQ ID NO: 979 or a fragment or variant thereof. As another
non-limiting example, the AADC polynucleotide comprises a sequence
which as 95% identity to any of SEQ ID NO: 979 or a fragment or
variant thereof. As another non-limiting example, the AADC
polynucleotide comprises a sequence which as 99% identity to any of
SEQ ID NO: 979 or a fragment or variant thereof.
[0279] In some embodiments, the coding region of the AADC
polynucleotide is 1440 nucleotides in length. Such an AADC
polynucleotide may be codon optimized over all or a portion of the
polynucleotide.
[0280] In some embodiments, the AADC polynucleotide comprises any
of SEQ ID NO: 979 or a fragment or variant thereof but lacking the
5' and/or 3' ITRs. Such a polynucleotide may be incorporated into a
plasmid or vector and utilized to express the encoded AADC
protein.
[0281] In one embodiment, the AADC polynucleotides may be produced
in insect cells (e.g., Sf9 cells).
[0282] In one embodiment, the AADC polynucleotides may be produced
using triple transfection.
[0283] In one embodiment, the AADC polynucleotide may comprise a
codon optimized open reading frame of an AADC mRNA, at least one
5'ITR and at least one 3'UTR where the one or more of the 5'ITRs
may be located at the 5'end of the promoter region and one or more
3' ITRs may be located at the 3' end of the poly(A) signal. The
AADC mRNA may comprise a promoter region, a 5'untranslated region
(UTR), a 3'UTR and a poly(A) signal. The promoter region may
include, but is not limited to, enhancer element, a promoter
element, a first exon region, a first intron region, a second
intron region and a second exon region. As a non-limiting example,
the enhancer element and the promoter element are derived from CMV.
As another non-limiting example, the first exon region is ie1 exon
1 or fragments thereof, the first intron region is ie1 intron 1 or
fragments thereof, the second intron region is hbBglobin intron 2
or fragments thereof and the second exon region is hbBglobin exon 3
or fragments thereof. As yet another non-limiting example, the
poly(A) signal is derived from human growth hormone.
[0284] In one embodiment, at least one element may be used with the
AADC polynucleotides described herein to enhance the transgene
target specificity and expression (See e.g., Powell et al. Viral
Expression Cassette Elements to Enhance Transgene Target
Specificity and Expression in Gene Therapy, 2015; the contents of
which are herein incorporated by reference in its entirety).
Non-limiting examples of elements to enhance the transgene target
specificity and expression include promoters, endogenous miRNAs,
post-transcriptional regulatory elements (PREs), polyadenylation
(PolyA) signal sequences and upstream enhancers (USEs), CMV
enhancers and introns.
[0285] In one embodiment, at least one element may be used with the
AADC polynucleotides described herein to enhance the transgene
target specificity and expression (See e.g., Powell et al. Viral
Expression Cassette Elements to Enhance Transgene Target
Specificity and Expression in Gene Therapy, 2015; the contents of
which are herein incorporated by reference in its entirety) such as
promoters.
[0286] In one embodiment, the AADC polynucleotide is encoded in a
plasmid or vector, which may be derived from an adeno-associated
virus (AAV). The AAV may comprise a capsid serotype such as, but
not limited to, PHP.B, PHP.A, AAV1, AAV2, AAV2G9, AAV3, AAV3a,
AAV3b, AAV3-3, AAV4, AAV4-4, AAV5, AAV6, AAV6.1, AAV6.2, AAV6.1.2,
AAV7, AAV7.2, AAV8, AAV9, AAV9.11, AAV9.13, AAV9.16, AAV9.24,
AAV9.45, AAV9.47, AAV9.61, AAV9.68, AAV9.84, AAV9.9, AAV10, AAV11,
AAV12, AAV16.3, AAV24.1, AAV27.3, AAV42.12, AAV42-lb, AAV42-2,
AAV42-3a, AAV42-3b, AAV42-4, AAV42-5a, AAV42-5b, AAV42-6b, AAV42-8,
AAV42-10, AAV42-11, AAV42-12, AAV42-13, AAV42-15, AAV42-aa,
AAV43-1, AAV43-12, AAV43-20, AAV43-21, AAV43-23, AAV43-25, AAV43-5,
AAV44.1, AAV44.2, AAV44.5, AAV223.1, AAV223.2, AAV223.4, AAV223.5,
AAV223.6, AAV223.7, AAV1-7/rh.48, AAV1-8/rh.49, AAV2-15/rh.62,
AAV2-3/rh.61, AAV2-4/rh.50, AAV2-5/rh.51, AAV3.1/hu.6, AAV3.1/hu.9,
AAV3-9/rh.52, AAV3-11/rh.53, AAV4-8/r11.64, AAV4-9/rh.54,
AAV4-19/rh.55, AAV5-3/rh.57, AAV5-22/rh.58, AAV7.3/hu.7,
AAV16.8/hu. 10, AAV16.12/hu.11, AAV29.3/bb.1, AAV29.5/bb.2,
AAV106.1/hu.37, AAV114.3/hu.40, AAV127.2/hu.41, AAV127.5/hu.42,
AAV128.3/hu.44, AAV130.4/hu.48, AAV145.1/hu.53, AAV145.5/hu.54,
AAV145.6/hu.55, AAV161.10/hu.60, AAV161.6/hu.61, AAV33.12/hu. 17,
AAV33.4/hu. 15, AAV33.8/hu. 16, AAV52/hu. 19, AAV52.1/hu.20,
AAV58.2/hu.25, AAVA3.3, AAVA3.4, AAVA3.5, AAVA3.7, AAVC1, AAVC2,
AAVC5, AAV-DJ, AAV-DJ8, AAVF3, AAVF5, AAVH2, AAVrh.72, AAVhu.8,
AAVrh.68, AAVrh.70, AAVpi.1, AAVpi.3, AAVpi.2, AAVrh.60, AAVrh.44,
AAVrh.65, AAVrh.55, AAVrh.47, AAVrh.69, AAVrh.45, AAVrh.59, AAVhu.
12, AAVH6, AAVLK03, AAVH-1/hu. 1, AAVH-5/hu.3, AAVLG-10/rh.40,
AAVLG-4/rh.38, AAVLG-9/hu.39, AAVN721-8/rh.43, AAVCh.5, AAVCh.5R1,
AAVcy.2, AAVcy.3, AAVcy.4, AAVcy.5, AAVCy.5R1, AAVCy.5R2,
AAVCy.5R3, AAVCy.5R4, AAVcy.6, AAVhu.1, AAVhu.2, AAVhu.3, AAVhu.4,
AAVhu.5, AAVhu.6, AAVhu.7, AAVhu.9, AAVhu.10, AAVhu.11, AAVhu.13,
AAVhu.15, AAVhu.16, AAVhu.17, AAVhu.18, AAVhu.20, AAVhu.21,
AAVhu.22, AAVhu.23.2, AAVhu.24, AAVhu.25, AAVhu.27, AAVhu.28,
AAVhu.29, AAVhu.29R, AAVhu.31, AAVhu.32, AAVhu.34, AAVhu.35,
AAVhu.37, AAVhu.39, AAVhu.40, AAVhu.41, AAVhu.42, AAVhu.43,
AAVhu.44, AAVhu.44R1, AAVhu.44R2, AAVhu.44R3, AAVhu.45, AAVhu.46,
AAVhu.47, AAVhu.48, AAVhu.48R1, AAVhu.48R2, AAVhu.48R3, AAVhu.49,
AAVhu.51, AAVhu.52, AAVhu.54, AAVhu.55, AAVhu.56, AAVhu.57,
AAVhu.58, AAVhu.60, AAVhu.61, AAVhu.63, AAVhu.64, AAVhu.66,
AAVhu.67, AAVhu.14/9, AAVhu.t 19, AAVrh.2, AAVrh.2R, AAVrh.8,
AAVrh.8R, AAVrh.10, AAVrh.12, AAVrh.13, AAVrh.13R, AAVrh.14,
AAVrh.17, AAVrh.18, AAVrh.19, AAVrh.20, AAVrh.21, AAVrh.22,
AAVrh.23, AAVrh.24, AAVrh.25, AAVrh.31, AAVrh.32, AAVrh.33,
AAVrh.34, AAVrh.35, AAVrh.36, AAVrh.37, AAVrh.37R2, AAVrh.38,
AAVrh.39, AAVrh.40, AAVrh.46, AAVrh.48, AAVrh.48.1, AAVrh.48.1.2,
AAVrh.48.2, AAVrh.49, AAVrh.51, AAVrh.52, AAVrh.53, AAVrh.54,
AAVrh.56, AAVrh.57, AAVrh.58, AAVrh.61, AAVrh.64, AAVrh.64R1,
AAVrh.64R2, AAVrh.67, AAVrh.73, AAVrh.74, AAVrh8R, AAVrh8R A586R
mutant, AAVrh8R R533A mutant, AAAV, BAAV, caprine AAV, bovine AAV,
AAVhE1.1, AAVhEr1.5, AAVhER1.14, AAVhEr1.8, AAVhEr1.16, AAVhEr1.18,
AAVhEr1.35, AAVhEr1.7, AAVhEr1.36, AAVhEr2.29, AAVhEr2.4,
AAVhEr2.16, AAVhEr2.30, AAVhEr2.31, AAVhEr2.36, AAVhER1.23,
AAVhEr3.1, AAV2.5T, AAV-PAEC, AAV-LK01, AAV-LK02, AAV-LK03,
AAV-LK04, AAV-LK05, AAV-LK06, AAV-LK07, AAV-LK08, AAV-LK09,
AAV-LK10, AAV-LK 11, AAV-LK12, AAV-LK13, AAV-LK14, AAV-LK 15,
AAV-LK16, AAV-LK17, AAV-LK18, AAV-LK19, AAV-PAEC2, AAV-PAEC4,
AAV-PAEC6, AAV-PAEC7, AAV-PAEC8, AAV-PAEC11, AAV-PAEC12,
AAV-2-pre-miRNA-101, AAV-8h, AAV-8b, AAV-h, AAV-b, AAV SM 10-2, AAV
Shuffle 100-1, AAV Shuffle 100-3, AAV Shuffle 100-7, AAV Shuffle
10-2, AAV Shuffle 10-6, AAV Shuffle 10-8, AAV Shuffle 100-2, AAV SM
10-1, AAV SM 10-8, AAV SM 100-3, AAV SM 100-10, BNP61 AAV, BNP62
AAV, BNP63 AAV, AAVrh.50, AAVrh.43, AAVrh.62, AAVrh.48, AAVhu.19,
AAVhu.11, AAVhu.53, AAV4-8/rh.64, AAVLG-9/hu.39, AAV54.5/hu.23,
AAV54.2/hu.22, AAV54.7/hu.24, AAV54.1/hu.21, AAV54.4R/hu.27,
AAV46.2/hu.28, AAV46.6/hu.29, AAV128.1/hu.43, true type AAV
(ttAAV), UPENN AAV 10, Japanese AAV 10 serotypes, AAV CBr-7.1, AAV
CBr-7.10, AAV CBr-7.2, AAV CBr-7.3, AAV CBr-7.4, AAV CBr-7.5, AAV
CBr-7.7, AAV CBr-7.8, AAV CBr-B7.3, AAV CBr-B7.4, AAV CBr-E1, AAV
CBr-E2, AAV CBr-E3, AAV CBr-E4, AAV CBr-E5, AAV CBr-e5, AAV CBr-E6,
AAV CBr-E7, AAV CBr-E8, AAV CHt-1, AAV CHt-2, AAV CHt-3, AAV
CHt-6.1, AAV CHt-6.10, AAV CHt-6.5, AAV CHt-6.6, AAV CHt-6.7, AAV
CHt-6.8, AAV CHt-P1, AAV CHt-P2, AAV CHt-P5, AAV CHt-P6, AAV
CHt-P8, AAV CHt-P9, AAV CKd-1, AAV CKd-10, AAV CKd-2, AAV CKd-3,
AAV CKd-4, AAV CKd-6, AAV CKd-7, AAV CKd-8, AAV CKd-B1, AAV CKd-B2,
AAV CKd-B3, AAV CKd-B4, AAV CKd-B5, AAV CKd-B6, AAV CKd-B7, AAV
CKd-B8, AAV CKd-H1, AAV CKd-H2, AAV CKd-H3, AAV CKd-H4, AAV CKd-H5,
AAV CKd-H6, AAV CKd-N3, AAV CKd-N4, AAV CKd-N9, AAV CLg-F1, AAV
CLg-F2, AAV CLg-F3, AAV CLg-F4, AAV CLg-F5, AAV CLg-F6, AAV CLg-F7,
AAV CLg-F8, AAV CLv-1, AAV CLv1-1, AAV Clv1-10, AAV CLv1-2, AAV
CLv-12, AAV CLv1-3, AAV CLv-13, AAV CLv1-4, AAV Clv1-7, AAV Clv1-8,
AAV Clv1-9, AAV CLv-2, AAV CLv-3, AAV CLv-4, AAV CLv-6, AAV CLv-8,
AAV CLv-D1, AAV CLv-D2, AAV CLv-D3, AAV CLv-D4, AAV CLv-D5, AAV
CLv-D6, AAV CLv-D7, AAV CLv-D8, AAV CLv-E1, AAV CLv-K1, AAV CLv-K3,
AAV CLv-K6, AAV CLv-L4, AAV CLv-L5, AAV CLv-L6, AAV CLv-M1, AAV
CLv-M11, AAV CLv-M2, AAV CLv-M5, AAV CLv-M6, AAV CLv-M7, AAV
CLv-M8, AAV CLv-M9, AAV CLv-R1, AAV CLv-R2, AAV CLv-R3, AAV CLv-R4,
AAV CLv-R5, AAV CLv-R6, AAV CLv-R7, AAV CLv-R8, AAV CLv-R9, AAV
CSp-1, AAV CSp-10, AAV CSp-11, AAV CSp-2, AAV CSp-3, AAV CSp-4, AAV
CSp-6, AAV CSp-7, AAV CSp-8, AAV CSp-8.10, AAV CSp-8.2, AAV
CSp-8.4, AAV CSp-8.5, AAV CSp-8.6, AAV CSp-8.7, AAV CSp-8.8, AAV
CSp-8.9, AAV CSp-9, AAV.hu.48R3, AAV.VR-355, AAV3B, AAV4, AAV5,
AAVF1/HSC1, AAVF11/HSC11, AAVF12/HSC12, AAVF13/HSC13, AAVF14/HSC14,
AAVF15/HSC15, AAVF16/HSC16, AAVF17/HSC17, AAVF2/HSC2, AAVF3/HSC3,
AAVF4/HSC4, AAVF5/HSC5, AAVF6/HSC6, AAVF7/HSC7, AAVF8/HSC8,
AAVF9/HSC9, PHP.B (AAV-PHP.B), PHP.A (AAV.PHP.A), G2B-26, G2B-13,
TH1.1-32, TH1.1-35, AAVPHP.B2, AAVPHP.B3, AAVPHP.N/PHP.B-DGT,
AAVPHP.B-EST, AAVPHP.B-GGT, AAVPHP.B-ATP, AAVPHP.B-ATT-T,
AAVPHP.B-DGT-T, AAVPHP.B-GGT-T, AAVPHP.B-SGS, AAVPHP.B-AQP,
AAVPHP.B-QQP, AAVPHP.B-SNP(3), AAVPHP.B-SNP, AAVPHP.B-QGT,
AAVPHP.B-NQT, AAVPHP.B-EGS, AAVPHP.B-SGN, AAVPHP.B-EGT,
AAVPHP.B-DST, AAVPHP.B-DST, AAVPHP.B-STP, AAVPHP.B-PQP,
AAVPHP.B-SQP, AAVPHP.B-QLP, AAVPHP.B-TMP, AAVPHP.B-TTP,
AAVPHP.S/G2A12, AAVG2A15/G2A3, AAVG2B4, and/or AAVG2B5, and
variants thereof.
II. Formulation and Delivery
Pharmaceutical Compositions
[0287] According to the present disclosure the AAV particles may be
prepared as pharmaceutical compositions. It will be understood that
such compositions necessarily comprise one or more active
ingredients and, most often, a pharmaceutically acceptable
excipient.
[0288] Relative amounts of the active ingredient (e.g. AAV
particle), a pharmaceutically acceptable excipient, and/or any
additional ingredients in a pharmaceutical composition in
accordance with the present disclosure may vary, depending upon the
identity, size, and/or condition of the subject being treated and
further depending upon the route by which the composition is to be
administered. For example, the composition may comprise between
0.1% and 99% (w/w) of the active ingredient. By way of example, the
composition may comprise between 0.1% and 100%, e.g., between 0.5
and 50%, between 1-30%, between 5-80%, at least 80% (w/w) active
ingredient. In some embodiments, the AAV particle pharmaceutical
compositions described herein may comprise at least one payload. As
a non-limiting example, the pharmaceutical compositions may contain
an AAV particle with 1, 2, 3, 4 or 5 payloads.
[0289] Although the descriptions of pharmaceutical compositions
provided herein are principally directed to pharmaceutical
compositions which are suitable for administration to humans, it
will be understood by the skilled artisan that such compositions
are generally suitable for administration to any other animal,
e.g., to non-human animals, e.g. non-human mammals.
[0290] Modification of pharmaceutical compositions suitable for
administration to humans in order to render the compositions
suitable for administration to various animals is well understood,
and the ordinarily skilled veterinary pharmacologist can design
and/or perform such modification with merely ordinary, if any,
experimentation. Subjects to which administration of the
pharmaceutical compositions is contemplated include, but are not
limited to, humans and/or other primates; mammals, including
commercially relevant mammals such as cattle, pigs, horses, sheep,
cats, dogs, mice, rats, birds, including commercially relevant
birds such as poultry, chickens, ducks, geese, and/or turkeys.
[0291] In some embodiments, compositions are administered to
humans, human patients or subjects.
Formulations
[0292] Formulations of the present disclosure can include, without
limitation, saline, liposomes, lipid nanoparticles, polymers,
peptides, proteins, cells transfected with AAV particles (e.g., for
transfer or transplantation into a subject) and combinations
thereof.
[0293] Formulations of the pharmaceutical compositions described
herein may be prepared by any method known or hereafter developed
in the art of pharmacology. As used herein the term "pharmaceutical
composition" refers to compositions comprising at least one active
ingredient and optionally one or more pharmaceutically acceptable
excipients.
[0294] In general, such preparatory methods include the step of
associating the active ingredient with an excipient and/or one or
more other accessory ingredients. As used herein, the phrase
"active ingredient" generally refers either to an AAV particle
carrying a payload region encoding the polypeptides of the
disclosure or to the end product encoded by a viral genome of by an
AAV particle as described herein.
[0295] Formulations of the AAV particles and pharmaceutical
compositions described herein may be prepared by any method known
or hereafter developed in the art of pharmacology. In general, such
preparatory methods include the step of bringing the active
ingredient into association with an excipient and/or one or more
other accessory ingredients, and then, if necessary and/or
desirable, dividing, shaping and/or packaging the product into a
desired single- or multi-dose unit.
[0296] A pharmaceutical composition in accordance with the present
disclosure may be prepared, packaged, and/or sold in bulk, as a
single unit dose, and/or as a plurality of single unit doses. As
used herein, a "unit dose" refers to a discrete amount of the
pharmaceutical composition comprising a predetermined amount of the
active ingredient. The amount of the active ingredient is generally
equal to the dosage of the active ingredient which would be
administered to a subject and/or a convenient fraction of such a
dosage such as, for example, one-half or one-third of such a
dosage.
[0297] In one embodiment, the AAV particles of the disclosure may
be formulated in PBS with 0.001% of pluronic acid (F-68) at a pH of
about 7.0.
[0298] In some embodiments, the AAV formulations described herein
may contain sufficient AAV particles for expression of at least one
expressed functional payload. As a non-limiting example, the AAV
particles may contain viral genomes encoding 1, 2, 3, 4 or 5
functional payloads.
[0299] According to the present disclosure AAV particles may be
formulated for CNS delivery. Agents that cross the brain blood
barrier may be used. For example, some cell penetrating peptides
that can target molecules to the brain blood barrier endothelium
may be used for formulation (e.g., Mathupala, Expert Opin Ther
Pat., 2009, 19, 137-140; the content of which is incorporated
herein by reference in its entirety).
Excipients and Diluents
[0300] The AAV particles of the disclosure can be formulated using
one or more excipients or diluents to (1) increase stability; (2)
increase cell transfection or transduction; (3) permit the
sustained or delayed release of the payload; (4) alter the
biodistribution (e.g., target the viral particle to specific
tissues or cell types); (5) increase the translation of encoded
protein; (6) alter the release profile of encoded protein and/or
(7) allow for regulatable expression of the payload of the
disclosure.
[0301] In some embodiments, a pharmaceutically acceptable excipient
may be at least 95%, at least 96%, at least 97%, at least 98%, at
least 99%, or 100% pure. In some embodiments, an excipient is
approved for use for humans and for veterinary use. In some
embodiments, an excipient may be approved by United States Food and
Drug Administration. In some embodiments, an excipient may be of
pharmaceutical grade. In some embodiments, an excipient may meet
the standards of the United States Pharmacopoeia (USP), the
European Pharmacopoeia (EP), the British Pharmacopoeia, and/or the
International Pharmacopoeia.
[0302] Excipients, as used herein, include, but are not limited to,
any and all solvents, dispersion media, diluents, or other liquid
vehicles, dispersion or suspension aids, surface active agents,
isotonic agents, thickening or emulsifying agents, preservatives,
and the like, as suited to the particular dosage form desired.
Various excipients for formulating pharmaceutical compositions and
techniques for preparing the composition are known in the art (see
Remington: The Science and Practice of Pharmacy, 21st Edition, A.
R. Gennaro, Lippincott, Williams & Wilkins, Baltimore, Md.,
2006; incorporated herein by reference in its entirety). The use of
a conventional excipient medium may be contemplated within the
scope of the present disclosure, except insofar as any conventional
excipient medium may be incompatible with a substance or its
derivatives, such as by producing any undesirable biological effect
or otherwise interacting in a deleterious manner with any other
component(s) of the pharmaceutical composition.
[0303] Exemplary diluents include, but are not limited to, calcium
carbonate, sodium carbonate, calcium phosphate, dicalcium
phosphate, calcium sulfate, calcium hydrogen phosphate, sodium
phosphate lactose, sucrose, cellulose, microcrystalline cellulose,
kaolin, mannitol, sorbitol, inositol, sodium chloride, dry starch,
cornstarch, powdered sugar, etc., and/or combinations thereof.
[0304] In one embodiment, the AAV particles may be formulated in a
hydrogel prior to administration. Hydrogels have a degree of
flexibility which is similar to natural tissue as a result of their
significant water content.
[0305] In another embodiment, a hydrogel may be administered to a
subject prior to the administration of an AAV particle formulation.
As a non-limiting example, the site of administration of the
hydrogel may be within 3 inches (e.g., within 2.9, 2.8, 2.7, 2.6,
2.5, 2.4, 2.3, 2.2., 2.1, 2.0, 1.9, 1.8, 1.7, 1.6, 1.5, 1.4, 1.3,
1.2, 1.1, 1.0, 0.9, 0.8, 0.7, 0.6, 0.5, 0.4, 0.3, 0.2, 0.1 or less
than 0.1 inches) of the site of administration of the AAV particle
formulation.
Inactive Ingredients
[0306] In some embodiments, AAV particle formulations may comprise
at least one inactive ingredient. As used herein, the term
"inactive ingredient" refers to one or more agents that do not
contribute to the activity of the active ingredient of the
pharmaceutical composition included in formulations. In some
embodiments, all, none or some of the inactive ingredients which
may be used in the formulations of the present disclosure may be
approved by the US Food and Drug Administration (FDA).
[0307] In one embodiment, the AAV particle pharmaceutical
compositions comprise at least one inactive ingredient such as, but
not limited to, 1,2,6-Hexanetriol;
1,2-Dimyristoyl-Sn-Glycero-3-(Phospho-S-(1-Glycerol));
1,2-Dimyristoyl-Sn-Glycero-3-Phosphocholine;
1,2-Dioleoyl-Sn-Glycero-3-Phosphocholine;
1,2-Dipalmitoyl-Sn-Glycero-3-(Phospho-Rac-(1-Glycerol));
1,2-Distearoyl-Sn-Glycero-3-(Phospho-Rac-(1-Glycerol));
1,2-Distearoyl-Sn-Glycero-3-Phosphocholine; 1-O-Tolylbiguanide;
2-Ethyl-1,6-Hexanediol; Acetic Acid; Acetic Acid, Glacial; Acetic
Anhydride; Acetone; Acetone Sodium Bisulfite; Acetylated Lanolin
Alcohols; Acetylated Monoglycerides; Acetylcysteine;
Acetyltryptophan, DL-; Acrylates Copolymer; Acrylic Acid-Isooctyl
Acrylate Copolymer; Acrylic Adhesive 788; Activated Charcoal;
Adcote 72A103; Adhesive Tape; Adipic Acid; Aerotex Resin 3730;
Alanine; Albumin Aggregated; Albumin Colloidal; Albumin Human;
Alcohol; Alcohol, Dehydrated; Alcohol, Denatured; Alcohol, Diluted;
Alfadex; Alginic Acid; Alkyl Ammonium Sulfonic Acid Betaine; Alkyl
Aryl Sodium Sulfonate; Allantoin; Allyl.Alpha.-Ionone; Almond Oil;
Alpha-Terpineol; Alpha-Tocopherol; Alpha-Tocopherol Acetate, D1-;
Alpha-Tocopherol, D1-; Aluminum Acetate; Aluminum Chlorhydroxy
Allantoinate; Aluminum Hydroxide; Aluminum Hydroxide-Sucrose,
Hydrated; Aluminum Hydroxide Gel; Aluminum Hydroxide Gel F 500;
Aluminum Hydroxide Gel F 5000; Aluminum Monostearate; Aluminum
Oxide; Aluminum Polyester; Aluminum Silicate; Aluminum Starch
Octenylsuccinate; Aluminum Stearate; Aluminum Subacetate; Aluminum
Sulfate Anhydrous; Amerchol C; Amerchol-Cab; Aminomethylpropanol;
Ammonia; Ammonia Solution; Ammonia Solution, Strong; Ammonium
Acetate; Ammonium Hydroxide; Ammonium Lauryl Sulfate; Ammonium
Nonoxynol-4 Sulfate; Ammonium Salt Of C-12-C-15 Linear Primary
Alcohol Ethoxylate; Ammonium Sulfate; Ammonyx; Amphoteric-2;
Amphoteric-9; Anethole; Anhydrous Citric Acid; Anhydrous Dextrose;
Anhydrous Lactose; Anhydrous Trisodium Citrate; Aniseed Oil; Anoxid
Sbn; Antifoam; Antipyrine; Apaflurane; Apricot Kernel Oil Peg-6
Esters; Aquaphor; Arginine; Arlacel; Ascorbic Acid; Ascorbyl
Palmitate; Aspartic Acid; Balsam Peru; Barium Sulfate; Beeswax;
Beeswax, Synthetic; Beheneth-10; Bentonite; Benzalkonium Chloride;
Benzenesulfonic Acid; Benzethonium Chloride; Benzododecinium
Bromide; Benzoic Acid; Benzyl Alcohol; Benzyl Benzoate; Benzyl
Chloride; Betadex; Bibapcitide; Bismuth Subgallate; Boric Acid;
Brocrinat; Butane; Butyl Alcohol; Butyl Ester Of Vinyl Methyl
Ether/Maleic Anhydride Copolymer (125000 Mw); Butyl Stearate;
Butylated Hydroxyanisole; Butylated Hydroxytoluene; Butylene
Glycol; Butylparaben; Butyric Acid; C20-40 Pareth-24; Caffeine;
Calcium; Calcium Carbonate; Calcium Chloride; Calcium Gluceptate;
Calcium Hydroxide; Calcium Lactate; Calcobutrol; Caldiamide Sodium;
Caloxetate Trisodium; Calteridol Calcium; Canada Balsam;
Caprylic/Capric Triglyceride; Caprylic/Capric/Stearic Triglyceride;
Captan; Captisol; Caramel; Carbomer 1342; Carbomer 1382; Carbomer
934; Carbomer 934p; Carbomer 940; Carbomer 941; Carbomer 980;
Carbomer 981; Carbomer Homopolymer Type B (Allyl Pentaerythritol
Crosslinked); Carbomer Homopolymer Type C (Allyl Pentaerythritol
Crosslinked); Carbon Dioxide; Carboxy Vinyl Copolymer;
Carboxymethylcellulose; Carboxymethylcellulose Sodium;
Carboxypolymethylene; Carrageenan; Carrageenan Salt; Castor Oil;
Cedar Leaf Oil; Cellulose; Cellulose, Microcrystalline;
Cerasynt-Se; Ceresin; Ceteareth-12; Ceteareth-15; Ceteareth-30;
Cetearyl Alcohol/Ceteareth-20; Cetearyl Ethylhexanoate; Ceteth-10;
Ceteth-2; Ceteth-20; Ceteth-23; Cetostearyl Alcohol; Cetrimonium
Chloride; Cetyl Alcohol; Cetyl Esters Wax; Cetyl Palmitate;
Cetylpyridinium Chloride; Chlorobutanol; Chlorobutanol Hemihydrate;
Chlorobutanol, Anhydrous; Chlorocresol; Chloroxylenol; Cholesterol;
Choleth; Choleth-24; Citrate; Citric Acid; Citric Acid Monohydrate;
Citric Acid, Hydrous; Cocamide Ether Sulfate; Cocamine Oxide; Coco
Betaine; Coco Diethanolamide; Coco Monoethanolamide; Cocoa Butter;
Coco-Glycerides; Coconut Oil; Coconut Oil, Hydrogenated; Coconut
Oil/Palm Kernel Oil Glycerides, Hydrogenated; Cocoyl
Caprylocaprate; Cola Nitida Seed Extract; Collagen; Coloring
Suspension; Corn Oil; Cottonseed Oil; Cream Base; Creatine;
Creatinine; Cresol; Croscarmellose Sodium; Crospovidone; Cupric
Sulfate; Cupric Sulfate Anhydrous; Cyclomethicone;
Cyclomethicone/Dimethicone Copolyol; Cysteine; Cysteine
Hydrochloride; Cysteine Hydrochloride Anhydrous; Cysteine, D1-;
D&C Red No. 28; D&C Red No. 33; D&C Red No. 36; D&C
Red No. 39; D&C Yellow No. 10; Dalfampridine; Daubert 1-5 Pestr
(Matte) 164z; Decyl Methyl Sulfoxide; Dehydag Wax Sx; Dehydroacetic
Acid; Dehymuls E; Denatonium Benzoate; Deoxycholic Acid; Dextran;
Dextran 40; Dextrin; Dextrose; Dextrose Monohydrate; Dextrose
Solution; Diatrizoic Acid; Diazolidinyl Urea; Dichlorobenzyl
Alcohol; Dichlorodifluoromethane; Dichlorotetrafluoroethane;
Diethanolamine; Diethyl Pyrocarbonate; Diethyl Sebacate; Diethylene
Glycol Monoethyl Ether; Diethylhexyl Phthalate; Dihydroxyaluminum
Aminoacetate; Diisopropanolamine; Diisopropyl Adipate; Diisopropyl
Dilinoleate; Dimethicone 350; Dimethicone Copolyol; Dimethicone
Mdx4-4210; Dimethicone Medical Fluid 360; Dimethyl Isosorbide;
Dimethyl Sulfoxide; Dimethylaminoethyl Methacrylate-Butyl
Methacrylate-Methyl Methacrylate Copolymer;
Dimethyldioctadecylammonium Bentonite; Dimethyl
siloxane/Methylvinylsiloxane Copolymer; Dinoseb Ammonium Salt;
Dipalmitoylphosphatidylglycerol, D1-; Dipropylene Glycol; Disodium
Cocoamphodiacetate; Disodium Laureth Sulfosuccinate; Disodium
Lauryl Sulfosuccinate; Disodium Sulfosalicylate; Disofenin;
Divinylbenzene Styrene Copolymer; Dmdm Hydantoin; Docosanol;
Docusate Sodium; Duro-Tak 280-2516; Duro-Tak 387-2516; Duro-Tak
80-1196; Duro-Tak 87-2070; Duro-Tak 87-2194; Duro-Tak 87-2287;
Duro-Tak 87-2296; Duro-Tak 87-2888; Duro-Tak 87-2979; Edetate
Calcium Disodium; Edetate Disodium; Edetate Disodium Anhydrous;
Edetate Sodium; Edetic Acid; Egg Phospholipids; Entsufon; Entsufon
Sodium; Epilactose; Epitetracycline Hydrochloride; Essence Bouquet
9200; Ethanolamine Hydrochloride; Ethyl Acetate; Ethyl Oleate;
Ethylcelluloses; Ethylene Glycol; Ethylene Vinyl Acetate Copolymer;
Ethylenediamine; Ethylenediamine Dihydrochloride;
Ethylene-Propylene Copolymer; Ethylene-Vinyl Acetate Copolymer (28%
Vinyl Acetate); Ethylene-Vinyl Acetate Copolymer (9% Vinylacetate);
Ethylhexyl Hydroxystearate; Ethylparaben; Eucalyptol; Exametazime;
Fat, Edible; Fat, Hard; Fatty Acid Esters; Fatty Acid
Pentaerythriol Ester; Fatty Acids; Fatty Alcohol Citrate; Fatty
Alcohols; Fd&C Blue No. 1; Fd&C Green No. 3; Fd&C Red
No. 4; Fd&C Red No. 40; Fd&C Yellow No. 10 (Delisted);
Fd&C Yellow No. 5; Fd&C Yellow No. 6; Ferric Chloride;
Ferric Oxide; Flavor 89-186; Flavor 89-259; Flavor Df-119; Flavor
Df-1530; Flavor Enhancer; Flavor Fig 827118; Flavor Raspberry
Pfc-8407; Flavor Rhodia Pharmaceutical No. Rf 451;
Fluorochlorohydrocarbons; Formaldehyde; Formaldehyde Solution;
Fractionated Coconut Oil; Fragrance 3949-5; Fragrance 520a;
Fragrance 6.007; Fragrance 91-122; Fragrance 9128-Y; Fragrance
93498g; Fragrance Balsam Pine No. 5124; Fragrance Bouquet 10328;
Fragrance Chemoderm 6401-B; Fragrance Chemoderm 6411; Fragrance
Cream No. 73457; Fragrance Cs-28197; Fragrance Felton 066m;
Fragrance Firmenich 47373; Fragrance Givaudan Ess 9090/1c;
Fragrance H-6540; Fragrance Herbal 10396; Fragrance Nj-1085;
Fragrance P O F1-147; Fragrance Pa 52805; Fragrance Pera Derm D;
Fragrance Rbd-9819; Fragrance Shaw Mudge U-7776; Fragrance Tf
044078; Fragrance Ungerer Honeysuckle K 2771; Fragrance Ungerer
N5195; Fructose; Gadolinium Oxide; Galactose; Gamma Cyclodextrin;
Gelatin; Gelatin, Crosslinked; Gelfoam Sponge; Gellan Gum (Low
Acyl); Gelva 737; Gentisic Acid; Gentisic Acid Ethanolamide;
Gluceptate Sodium; Gluceptate Sodium Dihydrate; Gluconolactone;
Glucuronic Acid; Glutamic Acid, D1-; Glutathione; Glycerin;
Glycerol Ester Of Hydrogenated Rosin; Glyceryl Citrate; Glyceryl
Isostearate; Glyceryl Laurate; Glyceryl Monostearate; Glyceryl
Oleate; Glyceryl Oleate/Propylene Glycol; Glyceryl Palmitate;
Glyceryl Ricinoleate; Glyceryl Stearate; Glyceryl
Stearate-Laureth-23; Glyceryl Stearate/Peg Stearate; Glyceryl
Stearate/Peg-100 Stearate; Glyceryl Stearate/Peg-40 Stearate;
Glyceryl Stearate-Stearamidoethyl Diethylamine; Glyceryl Trioleate;
Glycine; Glycine Hydrochloride; Glycol Distearate; Glycol Stearate;
Guanidine Hydrochloride; Guar Gum; Hair Conditioner (18n195-1m);
Heptane; Hetastarch; Hexylene Glycol; High Density Polyethylene;
Histidine; Human Albumin Microspheres; Hyaluronate Sodium;
Hydrocarbon; Hydrocarbon Gel, Plasticized; Hydrochloric Acid;
Hydrochloric Acid, Diluted; Hydrocortisone; Hydrogel Polymer;
Hydrogen Peroxide; Hydrogenated Castor Oil; Hydrogenated Palm Oil;
Hydrogenated Palm/Palm Kernel Oil Peg-6 Esters; Hydrogenated
Polybutene 635-690; Hydroxide Ion; Hydroxyethyl Cellulose;
Hydroxyethylpiperazine Ethane Sulfonic Acid; Hydroxymethyl
Cellulose; Hydroxyoctacosanyl Hydroxystearate; Hydroxypropyl
Cellulose; Hydroxypropyl Methylcellulose 2906;
Hydroxypropyl-Beta-cyclodextrin; Hypromellose 2208 (15000 Mpa.S);
Hypromellose 2910 (15000 Mpa.S); Hypromelloses; Imidurea; Iodine;
lodoxamic Acid; lofetamine Hydrochloride; Irish Moss Extract;
Isobutane; Isoceteth-20; Isoleucine; Isooctyl Acrylate; Isopropyl
Alcohol; Isopropyl Isostearate; Isopropyl Myristate; Isopropyl
Myristate-Myristyl Alcohol; Isopropyl Palmitate; Isopropyl
Stearate; Isostearic Acid; Isostearyl Alcohol; Isotonic Sodium
Chloride Solution; Jelene; Kaolin; Kathon Cg; Kathon Cg II;
Lactate; Lactic Acid; Lactic Acid, D1-; Lactic Acid, L-;
Lactobionic Acid; Lactose; Lactose Monohydrate; Lactose, Hydrous;
Laneth; Lanolin; Lanolin Alcohol-Mineral Oil; Lanolin Alcohols;
Lanolin Anhydrous; Lanolin Cholesterols; Lanolin Nonionic
Derivatives; Lanolin, Ethoxylated; Lanolin, Hydrogenated;
Lauralkonium Chloride; Lauramine Oxide; Laurdimonium Hydrolyzed
Animal Collagen; Laureth Sulfate; Laureth-2; Laureth-23; Laureth-4;
Lauric Diethanolamide; Lauric Myristic Diethanolamide; Lauroyl
Sarcosine; Lauryl Lactate; Lauryl Sulfate; Lavandula Angustifolia
Flowering Top; Lecithin; Lecithin Unbleached; Lecithin, Egg;
Lecithin, Hydrogenated; Lecithin, Hydrogenated Soy; Lecithin,
Soybean; Lemon Oil; Leucine; Levulinic Acid; Lidofenin; Light
Mineral Oil; Light Mineral Oil (85 Ssu); Limonene, (+/-)-; Lipocol
Sc-15; Lysine; Lysine Acetate; Lysine Monohydrate; Magnesium
Aluminum Silicate; Magnesium Aluminum Silicate Hydrate; Magnesium
Chloride; Magnesium Nitrate; Magnesium Stearate; Maleic Acid;
Mannitol; Maprofix; Mebrofenin; Medical Adhesive Modified S-15;
Medical Antiform A-F Emulsion; Medronate Disodium; Medronic Acid;
Meglumine; Menthol; Metacresol; Metaphosphoric Acid;
Methanesulfonic Acid; Methionine; Methyl Alcohol; Methyl
Gluceth-10; Methyl Gluceth-20; Methyl Gluceth-20 Sesquistearate;
Methyl Glucose Sesquistearate; Methyl Laurate; Methyl Pyrrolidone;
Methyl Salicylate; Methyl Stearate; Methylboronic Acid;
Methylcellulose (4000 Mpa.S); Methylcelluloses;
Methylchloroisothiazolinone; Methylene Blue; Methylisothiazolinone;
Methylparaben; Microcrystalline Wax; Mineral Oil; Mono And
Diglyceride; Monostearyl Citrate; Monothioglycerol; Multisterol
Extract; Myristyl Alcohol; Myristyl Lactate;
Myristyl-.Gamma.-Picolinium Chloride; N-(Carbamoyl-Methoxy
Peg-40)-1,2-Distearoyl-Cephalin Sodium; N,N-Dimethylacetamide;
Niacinamide; Nioxime; Nitric Acid; Nitrogen; Nonoxynol Iodine;
Nonoxynol-15; Nonoxynol-9; Norflurane; Oatmeal; Octadecene-1/Maleic
Acid Copolymer; Octanoic Acid; Octisalate; Octoxynol-1;
Octoxynol-40; Octoxynol-9; Octyldodecanol; Octylphenol
Polymethylene; Oleic Acid; Oleth-10/Oleth-5; Oleth-2; Oleth-20;
Oleyl Alcohol; Oleyl Oleate; Olive Oil; Oxidronate Disodium;
Oxyquinoline; Palm Kernel Oil; Palmitamine Oxide; Parabens;
Paraffin; Paraffin, White Soft; Parfum Creme 45/3; Peanut Oil;
Peanut Oil, Refined; Pectin; Peg 6-32 Stearate/Glycol Stearate; Peg
Vegetable Oil; Peg-100 Stearate; Peg-12 Glyceryl Laurate; Peg-120
Glyceryl Stearate; Peg-120 Methyl Glucose Dioleate; Peg-15
Cocamine; Peg-150 Distearate; Peg-2 Stearate; Peg-20 Sorbitan
Isostearate; Peg-22 Methyl Ether/Dodecyl Glycol Copolymer; Peg-25
Propylene Glycol Stearate; Peg-4 Dilaurate; Peg-4 Laurate; Peg-40
Castor Oil; Peg-40 Sorbitan Diisostearate; Peg-45/Dodecyl Glycol
Copolymer; Peg-5 Oleate; Peg-50 Stearate; Peg-54 Hydrogenated
Castor Oil; Peg-6 Isostearate; Peg-60 Castor Oil; Peg-60
Hydrogenated Castor Oil; Peg-7 Methyl Ether; Peg-75 Lanolin; Peg-8
Laurate; Peg-8 Stearate; Pegoxol 7 Stearate; Pentadecalactone;
Pentaerythritol Cocoate; Pentasodium Pentetate; Pentetate Calcium
Trisodium; Pentetic Acid; Peppermint Oil; Perflutren; Perfume
25677; Perfume Bouquet; Perfume E-1991; Perfume Gd 5604; Perfume
Tana 90/42 Scba; Perfume W-1952-1; Petrolatum; Petrolatum, White;
Petroleum Distillates; Phenol; Phenol, Liquefied; Phenonip;
Phenoxyethanol; Phenylalanine; Phenylethyl Alcohol; Phenylmercuric
Acetate; Phenylmercuric Nitrate; Phosphatidyl Glycerol, Egg;
Phospholipid; Phospholipid, Egg; Phospholipon 90g; Phosphoric Acid;
Pine Needle Oil (Pinus Sylvestris); Piperazine Hexahydrate;
Plastibase-50w; Polacrilin; Polidronium Chloride; Poloxamer 124;
Poloxamer 181; Poloxamer 182; Poloxamer 188; Poloxamer 237;
Poloxamer 407; Poly(Bis(P-Carboxyphenoxy)Propane Anhydride):
Sebacic Acid;
Poly(Dimethylsiloxane/Methylvinylsiloxane/Methylhydrogensiloxane)
Dimethylvinyl Or Dimethylhydroxy Or Trimethyl Endblocked;
Poly(D1-Lactic-Co-Glycolic Acid), (50:50;
Poly(D1-Lactic-Co-Glycolic Acid), Ethyl Ester Terminated, (50:50;
Polyacrylic Acid (250000 Mw); Polybutene (1400 Mw); Polycarbophil;
Polyester; Polyester Polyamine Copolymer; Polyester Rayon;
Polyethylene Glycol 1000; Polyethylene Glycol 1450; Polyethylene
Glycol 1500; Polyethylene Glycol 1540; Polyethylene Glycol 200;
Polyethylene Glycol 300; Polyethylene Glycol 300-1600; Polyethylene
Glycol 3350; Polyethylene Glycol 400; Polyethylene Glycol 4000;
Polyethylene Glycol 540; Polyethylene Glycol 600; Polyethylene
Glycol 6000; Polyethylene Glycol 8000; Polyethylene Glycol 900;
Polyethylene High Density Containing Ferric Oxide Black (<1%);
Polyethylene Low Density Containing Barium Sulfate (20-24%);
Polyethylene T; Polyethylene Terephthalates; Polyglactin;
Polyglyceryl-3 Oleate; Polyglyceryl-4 Oleate; Polyhydroxyethyl
Methacrylate; Polyisobutylene; Polyisobutylene (1100000 Mw);
Polyisobutylene (35000 Mw); Polyisobutylene 178-236;
Polyisobutylene 241-294; Polyisobutylene 35-39; Polyisobutylene Low
Molecular Weight; Polyisobutylene Medium Molecular Weight;
Polyisobutylene/Polybutene Adhesive; Polylactide; Polyols;
Polyoxyethylene-Polyoxypropylene 1800; Polyoxyethylene Alcohols;
Polyoxyethylene Fatty Acid Esters; Polyoxyethylene Propylene;
Polyoxyl 20 Cetostearyl Ether; Polyoxyl 35 Castor Oil; Polyoxyl 40
Hydrogenated Castor Oil; Polyoxyl 40 Stearate; Polyoxyl 400
Stearate; Polyoxyl 6 And Polyoxyl 32 Palmitostearate; Polyoxyl
Distearate; Polyoxyl Glyceryl Stearate; Polyoxyl Lanolin; Polyoxyl
Palmitate; Polyoxyl Stearate; Polypropylene; Polypropylene Glycol;
Polyquaternium-10; Polyquaternium-7 (70/30 Acrylamide/Dadmac;
Polysiloxane; Polysorbate 20; Polysorbate 40; Polysorbate 60;
Polysorbate 65; Polysorbate 80; Polyurethane; Polyvinyl Acetate;
Polyvinyl Alcohol; Polyvinyl Chloride; Polyvinyl Chloride-Polyvinyl
Acetate Copolymer; Polyvinylpyridine; Poppy Seed Oil; Potash;
Potassium Acetate; Potassium Alum; Potassium Bicarbonate; Potassium
Bisulfite; Potassium Chloride; Potassium Citrate; Potassium
Hydroxide; Potassium Metabisulfite; Potassium Phosphate, Dibasic;
Potassium Phosphate, Monobasic; Potassium Soap; Potassium Sorbate;
Povidone Acrylate Copolymer; Povidone Hydrogel; Povidone K17;
Povidone K25; Povidone K29/32; Povidone K30; Povidone K90; Povidone
K90f; Povidone/Eicosene Copolymer; Povidones; Ppg-12/Smdi
Copolymer; Ppg-15 Stearyl Ether; Ppg-20 Methyl Glucose Ether
Distearate; Ppg-26 Oleate; Product Wat; Proline; Promulgen D;
Promulgen G; Propane; Propellant A-46; Propyl Gallate; Propylene
Carbonate; Propylene Glycol; Propylene Glycol Diacetate; Propylene
Glycol Dicaprylate; Propylene Glycol Monolaurate; Propylene Glycol
Monopalmitostearate; Propylene Glycol Palmitostearate; Propylene
Glycol Ricinoleate; Propylene Glycol/Diazolidinyl
Urea/Methylparaben/Propylparben; Propylparaben; Protamine Sulfate;
Protein Hydrolysate; Pvm/Ma Copolymer; Quaternium-15; Quaternium-15
Cis-Form; Quaternium-52; Ra-2397; Ra-3011; Saccharin; Saccharin
Sodium; Saccharin Sodium Anhydrous; Safflower Oil; Sd Alcohol 3a;
Sd Alcohol 40; Sd Alcohol 40-2; Sd Alcohol 40b; Sepineo P 600;
Serine; Sesame Oil; Shea Butter; Silastic Brand Medical Grade
Tubing; Silastic Medical Adhesive, Silicone Type A; Silica, Dental;
Silicon; Silicon Dioxide; Silicon Dioxide, Colloidal; Silicone;
Silicone Adhesive 4102; Silicone Adhesive 4502; Silicone Adhesive
Bio-Psa Q7-4201; Silicone Adhesive Bio-Psa Q7-4301; Silicone
Emulsion; Silicone/Polyester Film Strip; Simethicone; Simethicone
Emulsion; Sipon Ls 20np; Soda Ash; Sodium Acetate; Sodium Acetate
Anhydrous; Sodium Alkyl Sulfate; Sodium Ascorbate; Sodium
Benzoate; Sodium Bicarbonate; Sodium Bisulfate; Sodium Bisulfite;
Sodium Borate; Sodium Borate Decahydrate; Sodium Carbonate; Sodium
Carbonate Decahydrate; Sodium Carbonate Monohydrate; Sodium
Cetostearyl Sulfate; Sodium Chlorate; Sodium Chloride; Sodium
Chloride Injection; Sodium Chloride Injection, Bacteriostatic;
Sodium Cholesteryl Sulfate; Sodium Citrate; Sodium Cocoyl
Sarcosinate; Sodium Desoxycholate; Sodium Dithionite; Sodium
Dodecylbenzenesulfonate; Sodium Formaldehyde Sulfoxylate; Sodium
Gluconate; Sodium Hydroxide; Sodium Hypochlorite; Sodium Iodide;
Sodium Lactate; Sodium Lactate, L-; Sodium Laureth-2 Sulfate;
Sodium Laureth-3 Sulfate; Sodium Laureth-5 Sulfate; Sodium Lauroyl
Sarcosinate; Sodium Lauryl Sulfate; Sodium Lauryl Sulfoacetate;
Sodium Metabisulfite; Sodium Nitrate; Sodium Phosphate; Sodium
Phosphate Dihydrate; Sodium Phosphate, Dibasic; Sodium Phosphate,
Dibasic, Anhydrous; Sodium Phosphate, Dibasic, Dihydrate; Sodium
Phosphate, Dibasic, Dodecahydrate; Sodium Phosphate, Dibasic,
Heptahydrate; Sodium Phosphate, Monobasic; Sodium Phosphate,
Monobasic, Anhydrous; Sodium Phosphate, Monobasic, Dihydrate;
Sodium Phosphate, Monobasic, Monohydrate; Sodium Polyacrylate
(2500000 Mw); Sodium Pyrophosphate; Sodium Pyrrolidone Carboxylate;
Sodium Starch Glycolate; Sodium Succinate Hexahydrate; Sodium
Sulfate; Sodium Sulfate Anhydrous; Sodium Sulfate Decahydrate;
Sodium Sulfite; Sodium Sulfosuccinated Undecyclenic
Monoalkylolamide; Sodium Tartrate; Sodium Thioglycolate; Sodium
Thiomalate; Sodium Thiosulfate; Sodium Thiosulfate Anhydrous;
Sodium Trimetaphosphate; Sodium Xylenesulfonate; Somay 44; Sorbic
Acid; Sorbitan; Sorbitan Isostearate; Sorbitan Monolaurate;
Sorbitan Monooleate; Sorbitan Monopalmitate; Sorbitan Monostearate;
Sorbitan Sesquioleate; Sorbitan Trioleate; Sorbitan Tristearate;
Sorbitol; Sorbitol Solution; Soybean Flour; Soybean Oil; Spearmint
Oil; Spermaceti; Squalane; Stabilized Oxychloro Complex; Stannous
2-Ethylhexanoate; Stannous Chloride; Stannous Chloride Anhydrous;
Stannous Fluoride; Stannous Tartrate; Starch; Starch 1500,
Pregelatinized; Starch, Corn; Stearalkonium Chloride; Stearalkonium
Hectorite/Propylene Carbonate; Stearamidoethyl Diethylamine;
Steareth-10; Steareth-100; Steareth-2; Steareth-20; Steareth-21;
Steareth-40; Stearic Acid; Stearic Diethanolamide;
Stearoxytrimethylsilane; Steartrimonium Hydrolyzed Animal Collagen;
Stearyl Alcohol; Sterile Water For Inhalation;
Styrene/Isoprene/Styrene Block Copolymer; Succimer; Succinic Acid;
Sucralose; Sucrose; Sucrose Distearate; Sucrose Polyesters;
Sulfacetamide Sodium; Sulfobutylether.Beta.-Cyclodextrin; Sulfur
Dioxide; Sulfuric Acid; Sulfurous Acid; Surfactol Qs; Tagatose, D-;
Talc; Tall Oil; Tallow Glycerides; Tartaric Acid; Tartaric Acid,
D1-; Tenox; Tenox-2; Tert-Butyl Alcohol; Tert-Butyl Hydroperoxide;
Tert-Butylhydroquinone;
Tetrakis(2-Methoxyisobutylisocyanide)Copper(I) Tetrafluoroborate;
Tetrapropyl Orthosilicate; Tetrofosmin; Theophylline; Thimerosal;
Threonine; Thymol; Tin; Titanium Dioxide; Tocopherol;
Tocophersolan; Total parenteral nutrition, lipid emulsion;
Triacetin; Tricaprylin; Trichloromonofluoromethane; Trideceth-10;
Triethanolamine Lauryl Sulfate; Trifluoroacetic Acid;
Triglycerides, Medium Chain; Trihydroxystearin; Trilaneth-4
Phosphate; Trilaureth-4 Phosphate; Trisodium Citrate Dihydrate;
Trisodium Hedta; Triton 720; Triton X-200; Trolamine; Tromantadine;
Tromethamine (TRIS); Tryptophan; Tyloxapol; Tyrosine; Undecylenic
Acid; Union 76 Amsco-Res 6038; Urea; Valine; Vegetable Oil;
Vegetable Oil Glyceride, Hydrogenated; Vegetable Oil, Hydrogenated;
Versetamide; Viscarin; Viscose/Cotton; Vitamin E; Wax, Emulsifying;
Wecobee Fs; White Ceresin Wax; White Wax; Xanthan Gum; Zinc; Zinc
Acetate; Zinc Carbonate; Zinc Chloride; and Zinc Oxide.
[0308] Pharmaceutical composition formulations of AAV particles
disclosed herein may include cations or anions. In one embodiment,
the formulations include metal cations such as, but not limited to,
Zn2+, Ca2+, Cu2+, Mn2+, Mg+ and combinations thereof. As a
non-limiting example, formulations may include polymers and
complexes with a metal cation (See e.g., U.S. Pat. Nos. 6,265,389
and 6,555,525, each of which is herein incorporated by reference in
its entirety). Formulations of the disclosure may also include one
or more pharmaceutically acceptable salts.
[0309] As used herein, "pharmaceutically acceptable salts" refers
to derivatives of the disclosed compounds wherein the parent
compound is modified by converting an existing acid or base moiety
to its salt form (e.g., by reacting the free base group with a
suitable organic acid). Examples of pharmaceutically acceptable
salts include, but are not limited to, mineral or organic acid
salts of basic residues such as amines; alkali or organic salts of
acidic residues such as carboxylic acids; and the like.
Representative acid addition salts include acetate, acetic acid,
adipate, alginate, ascorbate, aspartate, benzenesulfonate, benzene
sulfonic acid, benzoate, bisulfate, borate, butyrate, camphorate,
camphorsulfonate, citrate, cyclopentanepropionate, digluconate,
dodecylsulfate, ethanesulfonate, fumarate, glucoheptonate,
glycerophosphate, hemisulfate, heptonate, hexanoate, hydrobromide,
hydrochloride, hydroiodide, 2-hydroxy-ethanesulfonate,
lactobionate, lactate, laurate, lauryl sulfate, malate, maleate,
malonate, methanesulfonate, 2-naphthalenesulfonate, nicotinate,
nitrate, oleate, oxalate, palmitate, pamoate, pectinate,
persulfate, 3-phenylpropionate, phosphate, picrate, pivalate,
propionate, stearate, succinate, sulfate, tartrate, thiocyanate,
toluenesulfonate, undecanoate, valerate salts, and the like.
Representative alkali or alkaline earth metal salts include sodium,
lithium, potassium, calcium, magnesium, and the like, as well as
nontoxic ammonium, quaternary ammonium, and amine cations,
including, but not limited to ammonium, tetramethylammonium,
tetraethylammonium, methylamine, dimethylamine, trimethylamine,
triethylamine, ethylamine, and the like. The pharmaceutically
acceptable salts of the present disclosure include the conventional
non-toxic salts of the parent compound formed, for example, from
non-toxic inorganic or organic acids.
[0310] The pharmaceutically acceptable salts of the present
disclosure can be synthesized from the parent compound which
contains a basic or acidic moiety by conventional chemical methods.
Generally, such salts can be prepared by reacting the free acid or
base forms of these compounds with a stoichiometric amount of the
appropriate base or acid in water or in an organic solvent, or in a
mixture of the two; generally, nonaqueous media like ether, ethyl
acetate, ethanol, isopropanol, or acetonitrile are preferred. Lists
of suitable salts are found in Remington's Pharmaceutical Sciences,
17th ed., Mack Publishing Company, Easton, Pa., 1985, p. 1418,
Pharmaceutical Salts: Properties, Selection, and Use, P. H. Stahl
and C. G. Wermuth (eds.), Wiley-VCH, 2008, and Berge et al.,
Journal of Pharmaceutical Science, 66, 1-19 (1977); the content of
each of which is incorporated herein by reference in their
entirety.
[0311] The term "pharmaceutically acceptable solvate," as used
herein, means a compound of the disclosure wherein molecules of a
suitable solvent are incorporated in the crystal lattice. A
suitable solvent is physiologically tolerable at the dosage
administered. Solvates may be prepared by crystallization,
recrystallization, or precipitation from a solution that includes
organic solvents, water, or a mixture thereof. Examples of suitable
solvents are ethanol, water (for example, mono-, di-, and
tri-hydrates), N-methylpyrrolidinone (NMP), dimethyl sulfoxide
(DMSO), N,N'-dimethylformamide (DMF), N,N'-dimethylacetamide
(DMAC), 1,3-dimethyl-2-imidazolidinone (DMEU),
1,3-dimethyl-3,4,5,6-tetrahydro-2-(1H)-pyrimidinone (DMPU),
acetonitrile (ACN), propylene glycol, ethyl acetate, benzyl
alcohol, 2-pyrrolidone, benzyl benzoate, and the like. When water
is the solvent, the solvate is referred to as a "hydrate."
III. Administration and Dosing
Administration
[0312] In one embodiment, the AAV particle may be administered to a
subject (e.g., to the CNS of a subject) in a therapeutically
effective amount to reduce the symptoms of the disease of the
central nervous system (e.g., Parkinson's Disease) of a subject
(e.g., determined using a known evaluation method).
[0313] The AAV particles of the present disclosure may be
administered by any delivery route which results in a
therapeutically effective outcome. These include, but are not
limited to, enteral (into the intestine), gastroenteral, epidural
(into the dura mater), oral (by way of the mouth), transdermal,
intracerebral (into the cerebrum), intracerebroventricular (into
the cerebral ventricles), epicutaneous (application onto the skin),
intradermal, (into the skin itself), subcutaneous (under the skin),
nasal administration (through the nose), intravenous (into a vein),
intravenous bolus, intravenous drip, intra-arterial (into an
artery), intramuscular (into a muscle), intracardiac (into the
heart), intraosseous infusion (into the bone marrow), intrathecal
(into the spinal canal), intraparenchymal (into brain tissue),
intraperitoneal, (infusion or injection into the peritoneum),
intravesical infusion, intravitreal, (through the eye),
intracavernous injection (into a pathologic cavity) intracavitary
(into the base of the penis), intravaginal administration,
intrauterine, extra-amniotic administration, transdermal (diffusion
through the intact skin for systemic distribution), transmucosal
(diffusion through a mucous membrane), transvaginal, insufflation
(snorting), sublingual, sublabial, enema, eye drops (onto the
conjunctiva), or in ear drops, auricular (in or by way of the ear),
buccal (directed toward the cheek), conjunctival, cutaneous, dental
(to a tooth or teeth), electro-osmosis, endocervical, endosinusial,
endotracheal, extracorporeal, hemodialysis, infiltration,
interstitial, intra-abdominal, intra-amniotic, intra-articular,
intrabiliary, intrabronchial, intrabursal, intracartilaginous
(within a cartilage), intracaudal (within the cauda equine),
intracisternal (within the cisterna magna cerebellomedularis),
intracorneal (within the cornea), dental intracoronal,
intracoronary (within the coronary arteries), intracorporus
cavernosum (within the dilatable spaces of the corporus cavernosa
of the penis), intradiscal (within a disc), intraductal (within a
duct of a gland), intraduodenal (within the duodenum), intradural
(within or beneath the dura), intraepidermal (to the epidermis),
intraesophageal (to the esophagus), intragastric (within the
stomach), intragingival (within the gingivae), intraileal (within
the distal portion of the small intestine), intralesional (within
or introduced directly to a localized lesion), intraluminal (within
a lumen of a tube), intralymphatic (within the lymph),
intramedullary (within the marrow cavity of a bone), intrameningeal
(within the meninges), intramyocardial (within the myocardium),
intraocular (within the eye), intraovarian (within the ovary),
intrapericardial (within the pericardium), intrapleural (within the
pleura), intraprostatic (within the prostate gland), intrapulmonary
(within the lungs or its bronchi), intrasinal (within the nasal or
periorbital sinuses), intraspinal (within the vertebral column),
intrasynovial (within the synovial cavity of a joint),
intratendinous (within a tendon), intratesticular (within the
testicle), intrathecal (within the cerebrospinal fluid at any level
of the cerebrospinal axis), intrathoracic (within the thorax),
intratubular (within the tubules of an organ), intratumor (within a
tumor), intratympanic (within the aurus media), intravascular
(within a vessel or vessels), intraventricular (within a
ventricle), iontophoresis (by means of electric current where ions
of soluble salts migrate into the tissues of the body), irrigation
(to bathe or flush open wounds or body cavities), laryngeal
(directly upon the larynx), nasogastric (through the nose and into
the stomach), occlusive dressing technique (topical route
administration which is then covered by a dressing which occludes
the area), ophthalmic (to the external eye), oropharyngeal
(directly to the mouth and pharynx), parenteral, percutaneous,
periarticular, peridural, perineural, periodontal, rectal,
respiratory (within the respiratory tract by inhaling orally or
nasally for local or systemic effect), retrobulbar (behind the pons
or behind the eyeball), soft tissue, subarachnoid, subconjunctival,
submucosal, topical, transplacental (through or across the
placenta), transtracheal (through the wall of the trachea),
transtympanic (across or through the tympanic cavity), ureteral (to
the ureter), urethral (to the urethra), vaginal, caudal block,
diagnostic, nerve block, biliary perfusion, cardiac perfusion,
photopheresis and spinal.
[0314] In some embodiments, compositions may be administered in a
way which allows them to cross the blood-brain barrier, vascular
barrier, or other epithelial barrier. The AAV particles of the
present disclosure may be administered in any suitable form, either
as a liquid solution or suspension, as a solid form suitable for
liquid solution or suspension in a liquid solution. The AAV
particles may be formulated with any appropriate and
pharmaceutically acceptable excipient.
[0315] In one embodiment, the AAV particles of the present
disclosure may be delivered to a subject via a single route
administration.
[0316] In one embodiment, the AAV particles of the present
disclosure may be delivered to a subject via a multi-site route of
administration. A subject may be administered at 2, 3, 4, 5 or more
than 5 sites.
[0317] In one embodiment, a subject may be administered the AAV
particles of the present disclosure using a bolus infusion.
[0318] In one embodiment, a subject may be administered the AAV
particles of the present disclosure using sustained delivery over a
period of minutes, hours or days. The infusion rate may be changed
depending on the subject, distribution, formulation or another
delivery parameter.
[0319] In one embodiment, the AAV particles of the present
disclosure may be delivered by intramuscular delivery route. (See,
e.g., U.S. Pat. No. 6,506,379; the content of which is incorporated
herein by reference in its entirety). Non-limiting examples of
intramuscular administration include an intravenous injection or a
subcutaneous injection.
[0320] In one embodiment, the AAV particles of the present
disclosure may be delivered by oral administration. Non-limiting
examples of oral administration include a digestive tract
administration and a buccal administration.
[0321] In one embodiment, the AAV particles of the present
disclosure may be delivered by intraocular delivery route. A
non-limiting example of intraocular administration include an
intravitreal injection.
[0322] In one embodiment, the AAV particles of the present
disclosure may be delivered by intranasal delivery route.
Non-limiting examples of intranasal delivery include administration
of nasal drops or nasal sprays.
[0323] In some embodiments, the AAV particles that may be
administered to a subject by peripheral injections. Non-limiting
examples of peripheral injections include intraperitoneal,
intramuscular, intravenous, conjunctival or joint injection. It was
disclosed in the art that the peripheral administration of AAV
particles can be transported to the central nervous system, for
example, to the motor neurons (e.g., U. S. Patent Publication Nos.
20100240739; and 20100130594; the content of each of which is
incorporated herein by reference in their entirety).
[0324] In one embodiment, the AAV particles may be delivered by
injection into the CSF pathway. Non-limiting examples of delivery
to the CSF pathway include intrathecal and intracerebroventricular
administration.
[0325] In one embodiment, the AAV particles may be delivered by
systemic delivery. As a non-limiting example, the systemic delivery
may be by intravascular administration.
[0326] In one embodiment, the AAV particles of the present
disclosure may be administered to a subject by intracranial
delivery (See, e.g., U.S. Pat. No. 8,119,611; the content of which
is incorporated herein by reference in its entirety).
[0327] In some embodiments, the AAV particles of the present
disclosure may be administered by injection. As a non-limiting
example, the AAV particles of the present disclosure may be
administered to a subject by injection.
[0328] In some embodiments, the AAV particles of the present
disclosure may be administered by muscular injection. As a
non-limiting example, the AAV particles of the present disclosure
may be administered to a subject by muscular administration.
[0329] In some embodiments, the AAV particles of the present
disclosure may be administered by intramuscular administration. As
a non-limiting example, the AAV particles of the present disclosure
may be administered to a subject by intramuscular
administration.
[0330] In one embodiment, the AAV particles of the present
disclosure are administered to a subject and transduce muscle of a
subject. As a non-limiting example, the AAV particles are
administered by intramuscular administration.
[0331] In some embodiments, the AAV particles of the present
disclosure may be administered via intraparenchymal injection. As a
non-limiting example, the AAV particles of the present disclosure
may be administered to a subject by intraparenchymal
administration.
[0332] In some embodiments, the AAV particles of the present
disclosure may be administered by intravenous administration. As a
non-limiting example, the AAV particles of the present disclosure
may be administered to a subject by intravenous administration.
[0333] In one embodiment, the AAV particles of the present
disclosure may be administered via intravenous delivery.
[0334] In one embodiment, the AAV particles of the present
disclosure may be administered via a single dose intravenous
delivery. As a non-limiting example, the single dose intravenous
delivery may be a one-time treatment. In the context of diseases of
the central nervous system (e.g., Parkinson's Disease), the single
dose intravenous delivery can produce durable relief for subjects
with central nervous system (e.g., Parkinson's Disease) and/or
related symptoms. The relief may last for minutes such as, but not
limited to, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16,
17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33,
34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50,
51, 52, 53, 54, 55, 56, 57, 58, 59 minutes or more than 59 minutes;
hours such as, but not limited to, 1, 2, 1, 2, 3, 4, 5, 6, 7, 8, 9,
10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26,
27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43,
44, 45, 46, 47, 48, or more than 48 hours; days such as, but not
limited to, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16,
17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, or more
than 31 days; weeks such as, but not limited to, 1, 2, 3, 4, 5, 6,
7, 8, 9, 10, 11, 12, 13, 14, 15, 16, or more than 16 weeks; months
such as, but not limited to, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12,
13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, or more than 24
months; years such as, but not limited to, 1, 2, 3, 4, 5, 6, 7, 8,
9, 10, 11, 12, 13, 14, 15, or more than 15 years.
[0335] In one embodiment, the AAV particles of the present
disclosure may be administered via intravenous delivery to the DRG
nociceptive neurons.
[0336] In one embodiment, the AAV particles of the present
disclosure may be administered via a single dose intravenous
delivery to the DRG nociceptive neurons. As a non-limiting example,
the single dose intravenous delivery may be a one-time treatment.
In the context of diseases of the central nervous system (e.g.,
Parkinson's Disease), the single dose intravenous delivery can
produce durable relief for subjects with diseases of the central
nervous system (e.g., Parkinson's Disease) and/or related symptoms.
The relief may last for minutes such as, but not limited to, 1, 2,
3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20,
21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37,
38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54,
55, 56, 57, 58, 59 minutes or more than 59 minutes; hours such as,
but not limited to, 1, 2, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12,
13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29,
30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46,
47, 48, or more than 48 hours; days such as, but not limited to, 1,
2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20,
21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, or more than 31 days;
weeks such as, but not limited to, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10,
11, 12, 13, 14, 15, 16, or more than 16 weeks; months such as, but
not limited to, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15,
16, 17, 18, 19, 20, 21, 22, 23, 24, or more than 24 months; years
such as, but not limited to, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12,
13, 14, 15, or more than 15 years.
[0337] In some embodiments, the AAV particles of the present
disclosure may be administered by intrathecal injection. As a
non-limiting example, the AAV particles of the present disclosure
may be administered by intrathecal injection.
[0338] In one embodiment, the AAV particle may be administered to
the cisterna magna in a therapeutically effective amount to
transduce spinal cord motor neurons and/or astrocytes. As a
non-limiting example, the AAV particle may be administered
intrathecally.
[0339] In one embodiment, the AAV particle may be administered
using intrathecal infusion in a therapeutically effective amount to
transduce spinal cord motor neurons and/or astrocytes.
[0340] In some embodiments, the AAV particles of the present
disclosure may be administered via a single dose intrathecal
injection. As a non-limiting example, the single dose intrathecal
injection may be a one-time treatment. In the context of diseases
of the central nervous system (e.g., Parkinson's Disease), the
single dose intrathecal injection can produce durable relief for
subjects with diseases of the central nervous system (e.g.,
Parkinson's Disease) and/or related symptoms. The relief may last
for minutes such as, but not limited to, 1, 2, 3, 4, 5, 6, 7, 8, 9,
10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26,
27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43,
44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59
minutes or more than 59 minutes; hours such as, but not limited to,
1, 2, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17,
18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34,
35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, or more
than 48 hours; days such as, but not limited to, 1, 2, 3, 4, 5, 6,
7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23,
24, 25, 26, 27, 28, 29, 30, 31, or more than 31 days; weeks such
as, but not limited to, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13,
14, 15, 16, or more than 16 weeks; months such as, but not limited
to, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18,
19, 20, 21, 22, 23, 24, or more than 24 months; years such as, but
not limited to, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15,
or more than 15 years.
[0341] In some embodiments, the AAV particles of the present
disclosure may be administered via intrathecal injection to the DRG
nociceptive neurons.
[0342] In some embodiments, the AAV particles of the present
disclosure may be administered via a single dose intrathecal
injection to the DRG nociceptive neurons. As a non-limiting
example, the single dose intrathecal injection may be a one-time
treatment. In the context of diseases of the central nervous system
(e.g., Parkinson's Disease), the single dose intrathecal injection
can produce durable relief for subjects with diseases of the
central nervous system (e.g., Parkinson's Disease) and/or related
symptoms. The relief may last for minutes such as, but not limited
to, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18,
19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35,
36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52,
53, 54, 55, 56, 57, 58, 59 minutes or more than 59 minutes; hours
such as, but not limited to, 1, 2, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10,
11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27,
28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44,
45, 46, 47, 48, or more than 48 hours; days such as, but not
limited to, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16,
17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, or more
than 31 days; weeks such as, but not limited to, 1, 2, 3, 4, 5, 6,
7, 8, 9, 10, 11, 12, 13, 14, 15, 16, or more than 16 weeks; months
such as, but not limited to, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12,
13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, or more than 24
months; years such as, but not limited to, 1, 2, 3, 4, 5, 6, 7, 8,
9, 10, 11, 12, 13, 14, 15, or more than 15 years.
[0343] In one embodiment, the AAV particle described herein is
administered via intrathecal (IT) infusion at C1. The infusion may
be for 1, 2, 3, 4, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15 or more than
15 hours.
[0344] In some embodiments, the AAV particles of the present
disclosure may be administered by intraparenchymal injection. As a
non-limiting example, the AAV particles of the present disclosure
may be administered to a subject by intraparenchymal injection.
[0345] In one embodiment, the AAV particle may be administered to
the cisterna magna in a therapeutically effective amount to
transduce spinal cord motor neurons and/or astrocytes. As a
non-limiting example, the AAV particle may be administered
intraparenchymal injection.
[0346] In some embodiments, the AAV particles of the present
disclosure may be administered by intraparenchymal injection and
intrathecal injection. As a non-limiting example, the AAV particles
of the present disclosure may be administered via intraparenchymal
injection and intrathecal injection.
[0347] In some embodiments, the AAV particles of the present
disclosure may be administered by subcutaneous injection. As a
non-limiting example, the AAV particles In one embodiment, the AAV
particles of the present disclosure may be administered to a
subject by subcutaneous injection.
[0348] In some embodiments, the AAV particles of the present
disclosure may be administered topically. As a non-limiting
example, the AAV particles of the present disclosure may be
administered to a subject topically.
[0349] In one embodiment, the AAV particles may be delivered by
direct injection into the brain. As a non-limiting example, the
brain delivery may be by intrastriatal administration.
[0350] In one embodiment, the AAV particles of the present
disclosure may be administered via intrastriatal injection.
[0351] In one embodiment, the AAV particles of the present
disclosure may be administered via intrastriatal injection and
another route of administration described herein.
[0352] In one embodiment, the AAV particles may be delivered by
more than one route of administration. As non-limiting examples of
combination administrations, AAV particles may be delivered by
intrathecal and intracerebroventricular, or by intravenous and
intraparenchymal administration.
[0353] In one embodiment, the AAV particle may be administered to
the CNS in a therapeutically effective amount to improve function
and/or survival for a subject with diseases of the central nervous
system (e.g., Parkinson's Disease). As a non-limiting example, the
vector may be administered by direct infusion into the
striatum.
[0354] The AAV particle may be administered in a "therapeutically
effective" amount, i.e., an amount that is sufficient to alleviate
and/or prevent at least one symptom associated with the disease, or
provide improvement in the condition of the subject.
[0355] In one embodiment, the catheter may be located at more than
one site in the spine for multi-site delivery. The AAV particle may
be delivered in a continuous and/or bolus infusion. Each site of
delivery may be a different dosing regimen or the same dosing
regimen may be used for each site of delivery. As a non-limiting
example, the sites of delivery may be in the cervical and the
lumbar region. As another non-limiting example, the sites of
delivery may be in the cervical region. As another non-limiting
example, the sites of delivery may be in the lumbar region.
[0356] In one embodiment, a subject may be analyzed for spinal
anatomy and pathology prior to delivery of the AAV particle
described herein. As a non-limiting example, a subject with
scoliosis may have a different dosing regimen and/or catheter
location compared to a subject without scoliosis.
[0357] In one embodiment, the orientation of the spine of the
subject during delivery of the AAV particle may be vertical to the
ground.
[0358] In another embodiment, the orientation of the spine of the
subject during delivery of the AAV particle may be horizontal to
the ground.
[0359] In one embodiment, the spine of the subject may be at an
angle as compared to the ground during the delivery of the AAV
particle. The angle of the spine of the subject as compared to the
ground may be at least 10, 20, 30, 40, 50, 60, 70, 80, 90, 100,
110, 120, 130, 140, 150 or 180 degrees.
[0360] In one embodiment, the delivery method and duration is
chosen to provide broad transduction in the spinal cord. As a
non-limiting example, intrathecal delivery is used to provide broad
transduction along the rostral-caudal length of the spinal cord. As
another non-limiting example, multi-site infusions provide a more
uniform transduction along the rostral-caudal length of the spinal
cord. As yet another non-limiting example, prolonged infusions
provide a more uniform transduction along the rostral-caudal length
of the spinal cord.
[0361] In one embodiment, administration occurs by a posterior
(e.g., back of the head) surgical delivery approach to the putamen.
As a non-limiting example, the average putaminal coverage is 50%
with posterior delivery and the surgical time is less than 10
hours.
[0362] In one embodiment, administration occurs by a transfrontal
(e.g., top of the head) surgical delivery approach to the putamen.
As a non-limiting example, the average putaminal coverage less than
50% with posterior delivery and the surgical time is more than 10
hours.
Parenteral and Injectable Administration
[0363] In some embodiments, pharmaceutical compositions, AAV
particles of the present disclosure may be administered
parenterally. Liquid dosage forms for oral and parenteral
administration include, but are not limited to, pharmaceutically
acceptable emulsions, microemulsions, solutions, suspensions,
syrups, and/or elixirs. In addition to active ingredients, liquid
dosage forms may comprise inert diluents commonly used in the art
such as, for example, water or other solvents, solubilizing agents
and emulsifiers such as ethyl alcohol, isopropyl alcohol, ethyl
carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate,
propylene glycol, 1,3-butylene glycol, dimethylformamide, oils (in
particular, cottonseed, groundnut, corn, germ, olive, castor, and
sesame oils), glycerol, tetrahydrofurfuryl alcohol, polyethylene
glycols and fatty acid esters of sorbitan, and mixtures thereof.
Besides inert diluents, oral compositions can include adjuvants
such as wetting agents, emulsifying and suspending agents,
sweetening, flavoring, and/or perfuming agents. In certain
embodiments for parenteral administration, compositions are mixed
with solubilizing agents such as CREMOPHOR.RTM., alcohols, oils,
modified oils, glycols, polysorbates, cyclodextrins, polymers,
and/or combinations thereof. In other embodiments, surfactants are
included such as hydroxypropylcellulose.
[0364] Injectable preparations, for example, sterile injectable
aqueous or oleaginous suspensions may be formulated according to
the known art using suitable dispersing agents, wetting agents,
and/or suspending agents. Sterile injectable preparations may be
sterile injectable solutions, suspensions, and/or emulsions in
nontoxic parenterally acceptable diluents and/or solvents, for
example, as a solution in 1,3-butanediol. Among the acceptable
vehicles and solvents that may be employed are water, Ringer's
solution, U.S.P., and isotonic sodium chloride solution. Sterile,
fixed oils are conventionally employed as a solvent or suspending
medium. For this purpose, any bland fixed oil can be employed
including synthetic mono- or diglycerides. Fatty acids such as
oleic acid can be used in the preparation of injectables.
[0365] Injectable formulations may be sterilized, for example, by
filtration through a bacterial-retaining filter, and/or by
incorporating sterilizing agents in the form of sterile solid
compositions which can be dissolved or dispersed in sterile water
or other sterile injectable medium prior to use.
[0366] In order to prolong the effect of active ingredients, it is
often desirable to slow the absorption of active ingredients from
subcutaneous or intramuscular injections. This may be accomplished
by the use of liquid suspensions of crystalline or amorphous
material with poor water solubility. The rate of absorption of
active ingredients depends upon the rate of dissolution which, in
turn, may depend upon crystal size and crystalline form.
Alternatively, delayed absorption of a parenterally administered
drug form is accomplished by dissolving or suspending the drug in
an oil vehicle. Injectable depot forms are made by forming
microencapsule matrices of the drug in biodegradable polymers such
as polylactide-polyglycolide. Depending upon the ratio of drug to
polymer and the nature of the particular polymer employed, the rate
of drug release can be controlled. Examples of other biodegradable
polymers include poly(orthoesters) and poly(anhydrides). Depot
injectable formulations are prepared by entrapping the drug in
liposomes or microemulsions which are compatible with body
tissues.
Depot Administration
[0367] As described herein, in some embodiments, pharmaceutical
compositions, AAV particles of the present disclosure are
formulated in depots for extended release. Generally, specific
organs or tissues ("target tissues") are targeted for
administration.
[0368] In some aspects of the disclosure, pharmaceutical
compositions, AAV particles of the present disclosure are spatially
retained within or proximal to target tissues. Provided are methods
of providing pharmaceutical compositions, AAV particles, to target
tissues of mammalian subjects by contacting target tissues (which
comprise one or more target cells) with pharmaceutical
compositions, AAV particles, under conditions such that they are
substantially retained in target tissues, meaning that least 10,
20, 30, 40, 50, 60, 70, 80, 85, 90, 95, 96, 97, 98, 99, 99.9, 99.99
or greater than 99.99% of the composition is retained in the target
tissues. Advantageously, retention is determined by measuring the
amount of pharmaceutical compositions, AAV particles, that enter
one or more target cells. For example, at least 1%, 5%, 10%, 20%,
30%, 40%, 50%, 60%, 70%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%,
99.9%, 99.99% or greater than 99.99% of pharmaceutical
compositions, AAV particles, administered to subjects are present
intracellularly at a period of time following administration. For
example, intramuscular injection to mammalian subjects may be
performed using aqueous compositions comprising pharmaceutical
compositions, AAV particles of the present disclosure and one or
more transfection reagents, and retention is determined by
measuring the amount of pharmaceutical compositions, AAV particles,
present in muscle cells. Certain aspects of the disclosure are
directed to methods of providing pharmaceutical compositions, AAV
particles of the present disclosure to a target tissues of
mammalian subjects, by contacting target tissues (comprising one or
more target cells) with pharmaceutical compositions, AAV particles
under conditions such that they are substantially retained in such
target tissues.
[0369] Pharmaceutical compositions, AAV particles comprise enough
active ingredient such that the effect of interest is produced in
at least one target cell. In some embodiments, pharmaceutical
compositions, AAV particles generally comprise one or more cell
penetration agents, although "naked" formulations (such as without
cell penetration agents or other agents) are also contemplated,
with or without pharmaceutically acceptable carriers.
Delivery to the Central Nervous System
[0370] In one embodiment, delivery of the pharmaceutical
compositions comprising AAV particles to cells of the central
nervous system (e.g., parenchyma) comprises infusion of up to 1 mL.
In one embodiment, delivery of the pharmaceutical compositions
comprising AAV particles to cells of the central nervous system
(e.g., parenchyma) may comprise infusion of 0.001, 0.002, 0.003,
0.004, 0.005, 0.010, 0.015, 0.020, 0.025, 0.030, 0.040, 0.050,
0.060, 0.070, 0.080, 0.090, 0.1, 0.2, 0.3, 0.4, 0.5, 0.6, 0.7, 0.8,
or 0.9 mL.
[0371] In one embodiment, delivery of pharmaceutical composition
comprising AAV particles to cells of the central nervous system
(e.g., parenchyma) comprises infusion of between about 1 mL to
about 120 mL. In one embodiment, delivery of pharmaceutical
composition comprising AAV particles to cells of the central
nervous system (e.g., parenchyma) may comprise infusion of 0.1, 1,
1.1, 1.2, 1.3, 1.4, 1.5, 1.6, 1.7, 1.8, 1.9, 2, 3, 4, 5, 6, 7, 8,
9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25,
26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42,
43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59,
60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76,
77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93,
94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107,
108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 118, 119, or 120
mL. In one embodiment, delivery of AAV particles to cells of the
central nervous system (e.g., parenchyma) comprises infusion of at
least 3 mL. In one embodiment, delivery of AAV particles to cells
of the central nervous system (e.g., parenchyma) consists of
infusion of 3 mL. In one embodiment, delivery of AAV particles to
cells of the central nervous system (e.g., parenchyma) comprises
infusion of at least 10 mL. In one embodiment, delivery of AAV
particles to cells of the central nervous system (e.g., parenchyma)
consists of infusion of 10 mL.
[0372] In one embodiment, the volume of the pharmaceutical
composition comprising AAV particles delivered to the cells of the
central nervous system (e.g., parenchyma) of a subject is 50 ul,
100 ul, 200 ul, 300 ul, 400 ul, 500 ul, 600 ul, 700 ul, 800 ul, 900
ul, 1000 ul, 1100 ul, 1200 ul, 1300 ul, 1400 ul, 1500 ul, 1600 ul,
1700 ul, 1800 ul, 1900 ul, 2000 ul or more than 2000 ul.
[0373] In one embodiment, the volume of the pharmaceutical
composition comprising AAV particles delivered to a region in both
hemispheres of a subject brain is 50 ul, 100 ul, 200 ul, 300 ul,
400 ul, 500 ul, 600 ul, 700 ul, 800 ul, 900 ul, 1000 ul, 1100 ul,
1200 ul, 1300 ul, 1400 ul, 1500 ul, 1600 ul, 1700 ul, 1800 ul, 1900
ul, 2000 ul or more than 2000 ul. As a non-limiting example, the
volume delivered to a region in both hemispheres is 200 ul. As
another non-limiting example, the volume delivered to a region in
both hemispheres is 900 ul. As yet another non-limiting example,
the volume delivered to a region in both hemispheres is 1800
ul.
[0374] In one embodiment, the volume of the pharmaceutical
composition comprising AAV particles delivered to the putamen in
both hemispheres of a subject brain is 50 ul, 100 ul, 200 ul, 300
ul, 400 ul, 450 ul, 500 ul, 600 ul, 700 ul, 800 ul, 900 ul, 1000
ul, 1100 ul, 1200 ul, 1300 ul, 1400 ul, 1500 ul, 1600 ul, 1700 ul,
1800 ul, 1900 ul, 2000 ul or more than 2000 ul. As a non-limiting
example, the volume delivered to the putamen in both hemispheres is
100 ul. As another non-limiting example, the volume delivered to
the putamen in both hemispheres is 200 ul. As a non-limiting
example, the volume delivered to the putamen in both hemispheres is
300 ul. As another non-limiting example, the volume delivered to
the putamen in both hemispheres is 450 ul. As another non-limiting
example, the volume delivered to the putamen in both hemispheres is
900 ul. As yet another non-limiting example, the volume delivered
to the putamen both hemispheres is 1800 ul.
[0375] In one embodiment, the volume of the pharmaceutical
composition comprising AAV particles delivered to a subject is 900
ul to each putamen.
[0376] In one embodiment, the volume of the pharmaceutical
composition comprising AAV particles delivered to a subject is 450
ul to each putamen.
[0377] In one embodiment, the total volume delivered to a subject
may be split between one or more administration sites e.g., 1, 2,
3, 4, 5 or more than 5 sites. As a non-limiting example, the total
volume is split between administration to the left and right
putamen. As another non-limiting example, the total volume is split
between two sites of administration to each of the left and right
putamen.
[0378] In one embodiment, the pharmaceutical composition comprising
AAV particles is administered using a fenestrated needle.
Non-limiting examples of fenestrated needles are described in U.S.
Pat. Nos. 8,333,734, 7,135,010, 7,575,572, 7,699,852, 4,411,657,
6,890,319, 6,613,026, 6,726,659, 6,565,572, 6,520,949, 6,382,212,
5,848,996, 5,759,179, 5,674,267, 5,588,960, 5,484,401, 5,199,441,
5,012,818, 4,474,569, 3,766,907, 3,552,394, the contents of each of
which are herein incorporated by reference in its entirety.
[0379] In one embodiment, a composition comprises at least one
payload described herein and the payloads are components of a viral
genome packaged in an AAV particle. The percent (%) ratio of AAV
particles comprising the payload to the AAV particles without the
payload (also referred to herein as empty capsids) in the
composition may be 0:100, 1:99, 0:90, 15:85, 25:75, 30:70, 50:50,
70:30, 85:15, 90:10, 99:1 or 100:0. As a non-limiting example, the
percent ratio of AAV particles comprising the payload to empty
capsids is 50:50. As another non-limiting example, the percent
ratio of AAV particles comprising the payload to empty capsids is
70:30. As another non-limiting example, the percent ratio of AAV
particles comprising the payload to empty capsids is 85:15. As
another non-limiting example, the percent ratio of AAV particles
comprising the payload to empty capsids is 100:0.
[0380] In one embodiment, the composition described herein
comprises at least 1, 5, 10, 15, 20, 25, 30, 35, 40, 45, 50, 51,
52, 53, 54, 55, 60, 65, 70, 75, 80, 81, 82, 83, 84, 85, 86, 87, 88,
89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99 or greater than 99% AAV
particles comprising the payload. As a non-limiting example, the
composition comprises at least 50% AAV particles comprising the
payload. As another non-limiting example, the composition comprises
at least 52% AAV particles comprising the payload. As another
non-limiting example, the composition comprises at least 58% AAV
particles comprising the payload. As another non-limiting example,
the composition comprises at least 70% AAV particles comprising the
payload. As another non-limiting example, the composition comprises
at least 83% AAV particles comprising the payload. As another
non-limiting example, the composition comprises at least 85% AAV
particles comprising the payload. As another non-limiting example,
the composition comprises at least 99% AAV particles comprising the
payload. As another non-limiting example, the composition comprises
100% AAV particles comprising the payload.
[0381] In one embodiment, the composition described herein
comprises 1-10%, 10-20%, 30-40%, 50-60%, 50-70%, 50-80%, 50-90%,
50-99%, 50-100%, 60-70%, 60-80%, 60-90%, 60-99%, 60-100%, 70-80%,
70-90%, 70-99%, 70-100%, 80-85%, 80-90%, 80-95%, 80-99%, 80-100%,
90-95%, 90-99%, or 90-100% AAV particles comprising the payload. As
a non-limiting example, the composition described herein comprises
50-100% AAV particles comprising the payload. As another
non-limiting example, the composition described herein comprises
50-60% AAV particles comprising the payload. As another
non-limiting example, the composition described herein comprises
80-99% AAV particles comprising the payload. As another
non-limiting example, the composition described herein comprises
80-90% AAV particles comprising the payload. As a non-limiting
example, the composition described herein comprises 80-95% AAV
particles comprising the payload. As a non-limiting example, the
composition described herein comprises 80-85% AAV particles
comprising the payload.
[0382] In one embodiment, the composition described herein
comprises less than 1, 5, 10, 15, 20, 25, 30, 35, 40, 45, 50, 51,
52, 53, 54, 55, 60, 65, 70, 75, 80, 81, 82, 83, 84, 85, 86, 87, 88,
89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99 or 100% empty particles.
As a non-limiting example, the composition comprises less than 50%
empty particles. As a non-limiting example, the composition
comprises less than 45% empty particles. As a non-limiting example,
the composition comprises less than 40% empty particles. As a
non-limiting example, the composition comprises less than 35% empty
particles. As a non-limiting example, the composition comprises
less than 30% empty particles. As a non-limiting example, the
composition comprises less than 25% empty particles. As a
non-limiting example, the composition comprises less than 20% empty
particles. As a non-limiting example, the composition comprises
less than 15% empty particles. As a non-limiting example, the
composition comprises less than 10% empty particles. As a
non-limiting example, the composition comprises less than 5% empty
particles. As a non-limiting example, the composition comprises
less than 1% empty particles.
[0383] In the composition described herein comprises 1-10%, 10-20%,
30-40%, 50-60%, 50-70%, 50-80%, 50-90%, 50-99%, 50-100%, 60-70%,
60-80%, 60-90%, 60-99%, 60-100%, 70-80%, 70-90%, 70-99%, 70-100%,
80-85%, 80-90%, 80-95%, 80-99%, 80-100%, 90-95%, 90-99%, or 90-100%
empty particles. As a non-limiting example, the composition
described herein comprises 30-40% empty particles. As another
non-limiting example, the composition described herein comprises
30-50% empty particles. As another non-limiting example, the
composition described herein comprises 30-60% empty particles. As
another non-limiting example, the composition described herein
comprises 30-70% empty particles. As a non-limiting example, the
composition described herein comprises 30-80% empty particles. As a
non-limiting example, the composition described herein comprises
30-90% empty particles.
[0384] In one embodiment, the ratio of distribution volume in the
parenchyma of an area of a subject to the infusion volume of an
area of a subject may be 0.1, 0.2, 0.3, 0.4, 0.5, 0.6, 0.7, 0.8,
0.9, 1.0, 1.1, 1.2, 1.3, 1.4, 1.5, 1.6, 1.7, 1.8, 1.9, 2.0, 2.1,
2.2, 2.3, 2.4, 2.5, 2.6, 2.7, 2.8, 2.9, 3.0, 3.1, 3.2, 3.3, 3.4,
3.5, 3.6, 3.7, 3.8, 3.9, 4.0, 4.1, 4.2, 4.3, 4.4, 4.5, 4.6, 4.7,
4.8, 4.9, 5.0, 5.1, 5.2, 5.3, 5.4, 5.5, 5.6, 5.7, 5.8, 5.9, 6.0 or
more than 6.0. As a non-limiting example, the ratio of distribution
volume in the parenchyma to infusion volume was 1.6 in the caudate
nucleus. As a non-limiting example, the ratio of distribution
volume in the parenchyma to infusion volume was 3.1 in the putamen.
As a non-limiting example, the distribution of the AAV particles in
the putamen may be 2-3 times the volume infused.
[0385] In one embodiment, the effectiveness of the dose, route of
administration and/or volume of administration may be evaluated
using various methods described herein such as, but not limited to,
PET imaging, L-DOPA challenge test (e.g., see Forsayeth et al.
2006, Mol. Ther. 14(4): 571-577), UPDRS scores and patient diaries
(e.g., Hauser diary). As a non-limiting example, a subject may have
decreased dyskinesia or periods of decreased dyskinesia after
administration of the Pharmaceutical composition comprising AAV
particles. As another non-limiting example, a subject may have a
decrease in Parkinson's Disease related symptoms including limited
mobility and dyskinesia. As yet another non-limiting example, a
subject may show improvement in off time and motor fluctuations.
The improvement may be at least 10%, at least 20%, at least 30%, at
least 40%, at least 50%, at least 60%, at least 70%, at least 80%,
at least 90% or greater than 90%. The improvement may last for
minutes (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25, 30, 35,
40, 45, 50, 55 or more than 55), hours (e.g., 1, 2, 3, 4, 5, 6, 7,
8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23 or
more than 24), days (e.g., 1, 2, 3, 4, 5, 6 or more than 7), weeks
(1, 2, 3, 4, 5, 6, 7 or more than 7), months (1, 2, 3, 4, 5, 6, 7,
8, 9, 10, 11 or more than 11) or years (e.g., 1, 2, 3, 4, 5, 6, 7,
8, 9 or more than 9).
[0386] In one embodiment, the selection of subjects for
administration of the AAV particles described herein and/or the
effectiveness of the dose, route of administration and/or volume of
administration may be evaluated using imaging of the perivascular
spaces (PVS) which are also known as Virchow-Robin spaces. PVS
surround the arterioles and venules as they perforate brain
parenchyma and are filled with cerebrospinal fluid
(CSF)/interstitial fluid. PVS are common in the midbrain, BG, and
centrum semiovale. While not wishing to be bound by theory, PVS may
play a role in the normal clearance of metabolites and have been
associated with worse cognition and several disease states
including Parkinson's disease. PVS are usually normal in size but
they can increase in size in a number of disease states. Potter et
al. (Cerebrovasc Dis. 2015 January; 39(4): 224-231; the contents of
which are herein incorporated by reference in its entirety)
developed a grading method where they studied a full range of PVS
and rated basal ganglia, centrum semiovale and midbrain PVS. They
used the frequency and range of PVS used by Mac and Lullich et al.
(J Neurol Neurosurg Psychiatry. 2004 November; 75(11):1519-23; the
contents of which are herein incorporated by reference in its
entirety) and Potter et al. gave 5 ratings to basal ganglia and
centrum semiovale PVS: 0 (none), 1 (1-10), 2 (11-20), 3 (21-40) and
4 (>40) and 2 ratings to midbrain PVS: 0 (non visible) or 1
(visible). The user guide for the rating system by Potter et al.
can be found at:
www.sbirc.ed.ac.uk/documents/epvs-rating-scale-user-guide.pdf.
[0387] In one embodiment, the selection of subjects for
administration of the AAV particles described herein and/or the
effectiveness of the dose, route of administration and/or volume of
administration may be evaluated using positron emission tomography
(PET) measurements of neuroimaging biomarkers such as, but not
limited to [.sup.18F]FDOPA. Neuroimaging biomarkers such as
[.sup.18F]FDOPA may be used to identify affected individuals and/or
may be used to detect a nigrostriatal defect prior to the onset of
clinical manifestations. Further, PET-based criteria may be used to
categorize subjects based on their nigrostriatal neuronal integrity
(e.g., abnormal, normal or uncertain nigrostriatal neuronal
integrity) (Rachette et al. Am J Med Genet B Neuropsychiatr Genet.
2006 Apr. 5; 141B(3): 245-249; the contents of which are herein
incorporated by reference in its entirety).
[0388] In one embodiment, a subject who may be administered a dose
of the AAV particles described herein may have advanced PD and
still respond to levodopa therapy but the subject also experiences
medically refractory motor complications (e.g., sever motor
fluctuations and/or dyskinesias that occur during levodopa and
other dopaminergic therapies despite adjustments in and
optimization of medication). The subject may be healthy enough to
undergo a neurosurgical procedure which may be determined by
methods known in the art. As a non-limiting example, the subject
may meet the selection criteria for deep brain stimulation (DBS).
The subject may have idiopathic PD, younger than 69 years of age,
have pronounced responses to levodopa, have medication-refractory
symptoms (e.g., motor fluctuation and/or dyskinesia) and/or have
little or no cognitive dysfunction.
[0389] In one embodiment, a subject who may be administered a dose
of the AAV particles described herein may also suffer from dementia
or cognitive impairment.
[0390] In one embodiment, a subject who may be administered a dose
of the AAV particles described herein may have been previously
treated with the same or similar therapeutic. In another
embodiment, a subject may have been treated with a therapeutic
which has been shown to reduce the symptoms of Parkinson's
Disease.
[0391] In one embodiment, a subject who may be administered a dose
of the AAV particles described herein may have failed to derive
adequate benefit from standard medical therapy. As a non-limiting
example, the subject may not have responded to treatment. As
another non-limiting example, a subject may have residual
disability despite treatment.
[0392] In one embodiment, a subject who may be administered a dose
of the AAV particles described herein may undergo testing to
evaluate the levels of neurotransmitter analytes to determine the
effectiveness of the dose. As a non-limiting example, CSF
neurotransmitters, plasma AADC activity and/or urine VLA may be
analyzed.
[0393] In one embodiment, a subject who may be administered a dose
of the AAV particles described herein may be videotaped or recorded
in order to monitor the progress of the subject during the course
of treatment.
Delivery to the Putamen
[0394] In one embodiment, the AAV particles may be administered to
the right putamen and/or the left putamen. The administration may
be at one or more sites in the putamen such as, but not limited to,
2 sites, 3 sites, 4 sites or more than 4 sites. As a non-limiting
example, the AAV particles are delivered to 2 sites in the left
putamen and 2 sites in the right putamen.
[0395] In one embodiment, the administration of the formulation of
the AAV particles to a subject provides coverage of the putamen of
a subject (e.g., the left and/or right putamen). In one aspect, the
administration of the AAV particles may provide at least 8%, 9%,
10%, 13%, 14%, 15%, 19%, 20%, 21%, 22%, 23%, 24%, 25%, 26%, 27%,
28%, 29%, 30%, 31%, 32%, 33%, 34%, 35%, 36%, 37%, 38%, 39%, 40%,
41%, 42%, 43%, 44%, 45%, 46%, 47%, 48%, 49%, 50%, 51%, 52%, 53%,
54%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95% or more than 95%
to the left and/or right putamen of a subject. As a non-limiting
example, the coverage is at least 20%. As a non-limiting example,
the coverage is at least 40%. In another aspect, the administration
of the AAV particles may provide at least 8%, 9%, 10%, 13%, 14%,
15%, 19%, 20%, 21%, 22%, 23%, 24%, 25%, 26%, 27%, 28%, 29%, 30%,
31%, 32%, 33%, 34%, 35%, 36%, 37%, 38%, 39%, 40%, 41%, 42%, 43%,
44%, 45%, 46%, 47%, 48%, 49%, 50%, 51%, 52%, 53%, 54%, 55%, 60%,
65%, 70%, 75%, 80%, 85%, 90%, 95% or more than 95% coverage of the
surface area of the left and/or right putamen of a subject. As a
non-limiting example, the total coverage is at least 20%. As a
non-limiting example, the total coverage is at least 40%. In yet
another aspect, the administration of the AAV particles may provide
10-40%, 20-40%, 20-30%, 20-35%, 20-50%, 30-40%, 35-40%, 30-60%,
40-70%, 50-80% or 60-90% coverage to the left and/or right putamen
of a subject or to the total surface area of the left and/or right
putamen of a subject.
[0396] In one embodiment, the administration of the formulation of
the AAV particles to a subject provides coverage of the posterior
putamen of a subject (e.g., the left and/or right posterior
putamen). In one aspect, the administration of the AAV particles
may provide at least 10%, 15%, 20%, 21%, 22%, 23%, 24%, 25%, 26%,
27%, 28%, 29%, 30%, 31%, 32%, 33%, 34%, 35%, 36%, 37%, 38%, 39%,
40%, 41%, 42%, 43%, 44%, 45%, 46%, 47%, 48%, 49%, 50%,51%, 52%,
53%, 54%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95% or more than
95% to the left and/or right posterior putamen of a subject. As a
non-limiting example, the coverage is at least 20%. As a
non-limiting example, the coverage is at least 40%. In another
aspect, the administration of the AAV particles may provide at
least 10%, 15%, 20%, 21%, 22%, 23%, 24%, 25%, 26%, 27%, 28%, 29%,
30%, 31%, 32%, 33%, 34%, 35%, 36%, 37%, 38%, 39%, 40%, 41%, 42%,
43%, 44%, 45%, 46%, 47%, 48%, 49%, 50%, 51%, 52%, 53%, 54%, 55%,
60%, 65%, 70%, 75%, 80%, 85%, 90%, 95% or more than 95% coverage of
the surface area of the left and/or right posterior putamen of a
subject. As a non-limiting example, the total coverage is at least
20%. As a non-limiting example, the total coverage is at least 40%.
In yet another aspect, the administration of the AAV particles may
provide 10-40%, 20-50%, 30-60%, 40-70%, 50-80% or 60-90% coverage
to the left and/or right posterior putamen of a subject or to the
total surface area of the left and/or right putamen of a
subject.
[0397] In one embodiment, the AAV particles described herein may be
administered using acute bilateral placement of catheters into each
putamen. The placement may use magnetic resonance image
(MRI)-guided stereotactic neurosurgical techniques known in the art
or described herein. Additionally, a contrast agent such as, but
not limited to a gadolinium based contrast agent (e.g.,
PROHANCE.RTM.) may be used in the formulation to monitor and
confirm the distribution of the formulation.
[0398] In one embodiment, a subject may be administered the AAV
particles in a bilateral stereotactic CED-assisted step infusion
into the putamen (e.g., the post commissural putamen).
[0399] In one embodiment, a subject may be administered the AAV
particles of the present disclosure at a dose of
4.5.times.10.sup.12 vector genomes at a volume of 900 ul per
putamen.
[0400] In one embodiment, a subject may be administered the AAV
particles of the present disclosure at a dose of
1.5.times.10.sup.12 vector genomes at a volume of 900 ul per
putamen.
[0401] In one embodiment, a subject may be administered the AAV
particles of the present disclosure at a dose of
7.5.times.10.sup.11 vector genomes at a volume of 450 ul per
putamen.
[0402] In one embodiment, a subject may be administered the AAV
particles with a bilateral surgical infusion into at least one
putamen using a posterior (i.e., back of the head) surgical
delivery approach. The number of posterior bilateral surgical
infusions may be one or more such as, but not limited to, 1
infusion, 2 infusions, 3 infusions, 4 infusions or more than 4
infusions. As a non-limiting example, the AAV particles are
delivered in the left putamen with one posterior bilateral surgical
infusion. As a non-limiting example, the AAV particles are
delivered in the right putamen with one posterior bilateral
surgical infusion. As a non-limiting example, the AAV particles are
delivered in the left putamen with two posterior bilateral surgical
infusions. As a non-limiting example, the AAV particles are
delivered in the right putamen with two posterior bilateral
surgical infusions. As a non-limiting example, the AAV particles
are delivered in the right and left putamen with two posterior
bilateral surgical infusions. As a non-limiting example, the AAV
particles are delivered in the left putamen with three posterior
bilateral surgical infusions. As a non-limiting example, the AAV
particles are delivered in the right putamen with three posterior
bilateral surgical infusions. As a non-limiting example, the AAV
particles are delivered in the right and left putamen with three
posterior bilateral surgical infusions. As a non-limiting example,
the AAV particles are delivered in the left putamen with four
posterior bilateral surgical infusions. As a non-limiting example,
the AAV particles are delivered in the right putamen with four
posterior bilateral surgical infusions. As a non-limiting example,
the AAV particles are delivered in the right and left putamen with
four posterior bilateral surgical infusions.
[0403] In one embodiment, a subject may be administered the AAV
particles with a bilateral surgical infusion into at least one
putamen using a transfrontal (i.e., top of the head) surgical
delivery approach. The number of bilateral surgical infusions may
be two or more such as, but not limited to, 2 infusions, 3
infusions, 4 infusions or more than 4 infusions. As a non-limiting
example, the AAV particles are delivered in the left putamen with 2
posterior bilateral surgical infusions. As a non-limiting example,
the AAV particles are delivered in the right putamen with 2
posterior bilateral surgical infusions. As a non-limiting example,
the AAV particles are delivered in the left and right putamen with
2 posterior bilateral surgical infusions.
Delivery to the SNpc and VTA or STN
[0404] In one embodiment, a subject may be administered the AAV
particles of the present disclosure safely delivered to substantia
nigra pars compacta (SNpc) and ventral tegmental area (VTA) via
bilateral infusions, or alternatively, intrastriatally (into the
caudate nucleus and putamen), or into the subthalamic nucleus
(STN).
Delivery, Dose and Regimen
[0405] The present disclosure provides methods of administering AAV
particles in accordance with the disclosure to a subject in need
thereof. The pharmaceutical, diagnostic, or prophylactic AAV
particles and compositions of the present disclosure may be
administered to a subject using any amount and any route of
administration effective for preventing, treating, managing, or
diagnosing diseases, disorders and/or conditions. The exact amount
required will vary from subject to subject, depending on the
species, age, and general condition of the subject, the severity of
the disease, the particular composition, its mode of
administration, its mode of activity, and the like. The subject may
be a human, a mammal, or an animal. Compositions in accordance with
the disclosure are typically formulated in unit dosage form for
ease of administration and uniformity of dosage. It will be
understood, however, that the total daily usage of the compositions
of the present disclosure may be decided by the attending physician
within the scope of sound medical judgment. The specific
therapeutically effective, prophylactically effective, or
appropriate diagnostic dose level for any particular individual
will depend upon a variety of factors including the disorder being
treated and the severity of the disorder; the activity of the
specific payload employed; the specific composition employed; the
age, body weight, general health, sex and diet of the patient; the
time of administration, route of administration, and rate of
excretion of the specific AAV particle employed; the duration of
the treatment; drugs used in combination or coincidental with the
specific AAV particle employed; and like factors well known in the
medical arts.
[0406] In one embodiment, delivery of the AAV particles of the
present disclosure results in minimal serious adverse events (SAEs)
as a result of the delivery of the AAV particles.
[0407] In one embodiment, the AAV particle may be delivered a
multi-dose regimen. The multi-dose regimen may be 2, 3, 4, 5, 6, 7,
8, 9, 10 or more than 10 doses.
[0408] In one embodiment, the AAV particle may be delivered to a
subject via a multi-site route of administration. A subject may be
administered the AAV particle at 2, 3, 4, 5 or more than 5
sites.
Dosage Levels
[0409] In certain embodiments, AAV particle pharmaceutical
compositions in accordance with the present disclosure may be
administered at dosage levels sufficient to deliver from about
0.0001 mg/kg to about 100 mg/kg, from about 0.001 mg/kg to about
0.05 mg/kg, from about 0.005 mg/kg to about 0.05 mg/kg, from about
0.001 mg/kg to about 0.005 mg/kg, from about 0.05 mg/kg to about
0.5 mg/kg, from about 0.01 mg/kg to about 50 mg/kg, from about 0.1
mg/kg to about 40 mg/kg, from about 0.5 mg/kg to about 30 mg/kg,
from about 0.01 mg/kg to about 10 mg/kg, from about 0.1 mg/kg to
about 10 mg/kg, or from about 1 mg/kg to about 25 mg/kg, of subject
body weight per day, one or more times a day, to obtain the desired
therapeutic, diagnostic, or prophylactic, effect. It will be
understood that the above dosing concentrations may be converted to
vg or viral genomes per kg or into total viral genomes administered
by one of skill in the art.
[0410] In certain embodiments, AAV particle pharmaceutical
compositions in accordance with the present disclosure may be
administered at about 10 to about 600 .mu.l/site, 50 to about 500
.mu.l/site, 100 to about 400 .mu.l/site, 120 to about 300
.mu.l/site, 140 to about 200 .mu.l/site, about 160 .mu.l/site. As
non-limiting examples, AAV particles may be administered at 50
.mu.l/site and/or 150 .mu.l/site.
[0411] In one embodiment, delivery of the compositions in
accordance with the present disclosure to cells comprises a rate of
delivery defined by [VG/hour=mL/hour*VG/mL] wherein VG is viral
genomes, VG/mL is composition concentration, and mL/hour is rate of
prolonged delivery.
[0412] In one embodiment, delivery of compositions in accordance
with the present disclosure to cells may comprise a total
concentration per subject between about 1.times.10.sup.6 VG and
about 1.times.10.sup.16 VG. In some embodiments, delivery may
comprise a composition concentration of about 1.times.10.sup.6,
2.times.10.sup.6, 3.times.10.sup.6, 4.times.10.sup.6,
5.times.10.sup.6, 6.times.10.sup.6, 7.times.10.sup.6,
8.times.10.sup.6, 9.times.10.sup.6, 1.times.10.sup.7,
2.times.10.sup.7, 3.times.10.sup.7, 4.times.10.sup.7,
5.times.10.sup.7, 6.times.10.sup.7, 7.times.10.sup.7,
8.times.10.sup.7, 9.times.10.sup.7, 1.times.10.sup.8,
2.times.10.sup.8, 3.times.10.sup.8, 4.times.10.sup.8,
5.times.10.sup.8, 6.times.10.sup.8, 7.times.10.sup.8,
8.times.10.sup.8, 9.times.10.sup.8, 1.times.10.sup.9,
2.times.10.sup.9, 3.times.10.sup.9, 4.times.10.sup.9,
5.times.10.sup.9, 6.times.10.sup.9, 7.times.10.sup.9,
8.times.10.sup.9, 9.times.10.sup.9, 1.times.10.sup.10,
2.times.10.sup.0, 3.times.10.sup.0, 4.times.10.sup.0,
5.times.10.sup.0, 6.times.10.sup.0, 7.times.10.sup.0,
8.times.10.sup.0, 9.times.10.sup.10, 1.times.10.sup.11,
2.times.10.sup.1, 2.1.times.10.sup.11, 2.2.times.10.sup.11,
2.3.times.10.sup.11, 2.4.times.10.sup.11, 2.5.times.10.sup.11,
2.6.times.10.sup.11, 2.7.times.10.sup.11, 2.8.times.10.sup.11,
2.9.times.10.sup.11, 3.times.10.sup.1, 4.times.10.sup.1,
5.times.10.sup.1, 6.times.10.sup.1, 7.times.10.sup.1,
7.1.times.10.sup.11, 7.2.times.10.sup.11, 7.3.times.10.sup.11,
7.4.times.10.sup.11, 7.5.times.10.sup.11, 7.6.times.10.sup.11,
7.7.times.10.sup.11, 7.8.times.10.sup.11, 7.9.times.10.sup.11,
8.times.10.sup.11, 9.times.10.sup.11, 1.times.10.sup.12,
1.1.times.10.sup.12, 1.2.times.10.sup.12, 1.3.times.10.sup.12,
1.4.times.10.sup.12, 1.5.times.10.sup.12, 1.6.times.10.sup.12,
1.7.times.10.sup.12, 1.8.times.10.sup.12, 1.9.times.10.sup.12,
2.times.10.sup.12, 3.times.10.sup.12, 4.times.10.sup.12,
4.1.times.10.sup.12, 4.2.times.10.sup.12, 4.3.times.10.sup.12,
4.4.times.10.sup.12, 4.5.times.10.sup.12, 4.6.times.10.sup.12,
4.7.times.10.sup.12, 4.8.times.10.sup.12, 4.9.times.10.sup.12,
5.times.10.sup.12, 6.times.10.sup.12, 7.times.10.sup.12,
8.times.10.sup.12, 8.1.times.10.sup.12, 8.2.times.10.sup.12,
8.3.times.10.sup.12, 8.4.times.10.sup.12, 8.5.times.10.sup.12,
8.6.times.10.sup.12, 8.7.times.10.sup.12, 8.8.times.10.sup.12,
8.9.times.10.sup.12, 9.times.10.sup.12, 1.times.10.sup.13,
2.times.10.sup.13, 3.times.10.sup.13, 4.times.10.sup.13,
5.times.10.sup.13, 6.times.10.sup.13, 6.7.times.10.sup.13,
7.times.10.sup.13, 8.times.10.sup.13, 9.times.10.sup.13,
1.times.10.sup.14, 2.times.10.sup.14, 3.times.10.sup.14,
4.times.10.sup.14, 5.times.10.sup.14, 6.times.10.sup.14,
7.times.10.sup.14, 8.times.10.sup.14, 9.times.10.sup.14,
1.times.10.sup.15, 2.times.10.sup.15, 3.times.10.sup.15,
4.times.10.sup.15, 5.times.10.sup.15, 6.times.10.sup.15,
7.times.10.sup.15, 8.times.10.sup.15, 9.times.10.sup.15, or
1.times.10.sup.16 VG/subject. In one embodiment, the concentration
of the AAV particle in the composition is 1.times.10.sup.13
VG/subject. In one embodiment, the concentration of the AAV
particle in the composition is 3.times.10.sup.12 VG/subject. As a
non-limiting example, the composition administered to the subject
has a concentration of about 3.times.10.sup.11 VG/subject. As a
non-limiting example, the composition administered to the subject
has a concentration of about 9.times.10.sup.11 VG/subject. In one
embodiment, the concentration of the AAV particle in the
composition is 2.3.times.10.sup.11 VG/subject. In one embodiment,
the concentration of the AAV particle in the composition is
7.2.times.10.sup.11 VG/subject. In one embodiment, the
concentration of the AAV particle in the composition is
7.5.times.10.sup.11 VG/subject. In one embodiment, the
concentration of the AAV particle in the composition is
1.4.times.10.sup.12 VG/subject. In one embodiment, the
concentration of the AAV particle in the composition is
4.8.times.10.sup.12 VG/subject. In one embodiment, the
concentration of the AAV particle in the composition is
8.8.times.10.sup.12 VG/subject. In one embodiment, the
concentration of the AAV particle in the composition is
2.3.times.10.sup.12 VG/subject.
[0413] In one embodiment, delivery of compositions in accordance
with the present disclosure to cells may comprise a total
concentration per subject between about 1.times.10.sup.6 VG/kg and
about 1.times.10.sup.16 VG/kg. In some embodiments, delivery may
comprise a composition concentration of about 1.times.10.sup.6,
2.times.10.sup.6, 3.times.10.sup.6, 4.times.10.sup.6,
5.times.10.sup.6, 6.times.10.sup.6, 7.times.10.sup.6,
8.times.10.sup.6, 9.times.10.sup.6, 1.times.10.sup.7,
2.times.10.sup.7, 3.times.10.sup.7, 4.times.10.sup.7,
5.times.10.sup.7, 6.times.10.sup.7, 7.times.10.sup.7,
8.times.10.sup.7, 9.times.10.sup.7, 1.times.10, 2.times.10,
3.times.10.sup.8, 4.times.10.sup.8, 5.times.10, 6.times.10,
7.times.10, 8.times.10, 9.times.10.sup.8, 1.times.10.sup.9,
2.times.10.sup.9, 3.times.10.sup.9, 4.times.10.sup.9,
5.times.10.sup.9, 6.times.10.sup.9, 7.times.10.sup.9,
8.times.10.sup.9, 9.times.10.sup.9, 1.times.10.sup.10,
2.times.10.sup.10, 3.times.10.sup.10, 4.times.10.sup.10,
5.times.10.sup.10, 6.times.10.sup.10, 7.times.10.sup.10,
8.times.10.sup.10, 9.times.10.sup.10, 1.times.10.sup.11,
2.times.10.sup.11, 2.1.times.10.sup.11, 2.2.times.10.sup.11,
2.3.times.10.sup.11, 2.4.times.10.sup.11, 2.5.times.10.sup.11,
2.6.times.10.sup.11, 2.7.times.10.sup.11, 2.8.times.10.sup.11,
2.9.times.10.sup.11, 3.times.10.sup.11, 4.times.10.sup.11,
5.times.10.sup.11, 6.times.10.sup.11, 7.times.10.sup.11,
7.1.times.10.sup.11, 7.2.times.10.sup.11, 7.3.times.10.sup.11,
7.4.times.10.sup.11, 7.5.times.10.sup.11, 7.6.times.10.sup.11,
7.7.times.10.sup.11, 7.8.times.10.sup.11, 7.9.times.10.sup.11,
8.times.10.sup.11, 9.times.10.sup.11, 1.times.10.sup.12,
1.1.times.10.sup.12, 1.2.times.10.sup.12, 1.3.times.10.sup.12,
1.4.times.10.sup.12, 1.5.times.10.sup.12, 1.6.times.10.sup.12,
1.7.times.10.sup.12, 1.8.times.10.sup.12, 1.9.times.10.sup.12,
2.times.10.sup.12, 3.times.10.sup.12, 4.times.10.sup.12,
4.1.times.10.sup.12, 4.2.times.10.sup.12, 4.3.times.10.sup.12,
4.4.times.10.sup.12, 4.5.times.10.sup.12, 4.6.times.10.sup.12,
4.7.times.10.sup.12, 4.8.times.10.sup.12, 4.9.times.10.sup.12,
5.times.10.sup.12, 6.times.10.sup.12, 7.times.10.sup.12
8.times.10.sup.12, 8.1.times.10.sup.12, 8.2.times.10.sup.12,
8.3.times.10.sup.12, 8.4.times.10.sup.12, 8.5.times.10.sup.12,
8.6.times.10.sup.12, 8.7.times.10.sup.12, 8.8.times.10.sup.12,
8.9.times.10.sup.12, 9.times.10.sup.12, 1.times.10.sup.13,
2.times.10.sup.13, 3.times.10.sup.13, 4.times.10.sup.13,
5.times.10.sup.13, 6.times.10.sup.13, 6.7.times.10.sup.13,
7.times.10.sup.13, 8.times.10.sup.13, 9.times.10.sup.13,
1.times.10.sup.14, 2.times.10.sup.14, 3.times.10.sup.14,
4.times.10.sup.14, 5.times.10.sup.14, 6.times.10.sup.14,
7.times.10.sup.14, 8.times.10.sup.14, 9.times.10.sup.14,
1.times.10.sup.15, 2.times.10.sup.15, 3.times.10.sup.15,
4.times.10.sup.15, 5.times.10.sup.15, 6.times.10.sup.15,
7.times.10.sup.15, 8.times.10.sup.15, 9.times.10.sup.15, or
1.times.10.sup.16 VG/kg.
[0414] In one embodiment, delivery of AAV particles to cells of the
central nervous system (e.g., parenchyma) may comprise a total dose
between about 1.times.10.sup.6 VG and about 1.times.10.sup.16 VG.
In some embodiments, delivery may comprise a total dose of about
1.times.10.sup.6, 2.times.10.sup.6, 3.times.10.sup.6,
4.times.10.sup.6, 5.times.10.sup.6, 6.times.10.sup.6,
7.times.10.sup.6, 8.times.10.sup.6, 9.times.10.sup.6,
1.times.10.sup.7, 2.times.10.sup.7, 3.times.10.sup.7,
4.times.10.sup.7, 5.times.10.sup.7, 6.times.10.sup.7,
7.times.10.sup.7, 8.times.10.sup.7, 9.times.10.sup.7,
1.times.10.sup.8, 2.times.10.sup.8, 3.times.10.sup.8,
4.times.10.sup.8, 5.times.10, 6.times.10, 7.times.10, 8.times.10,
9.times.10.sup.8, 1.times.10.sup.9, 2.times.10.sup.9,
3.times.10.sup.9, 4.times.10.sup.9, 5.times.10.sup.9,
6.times.10.sup.9, 7.times.10.sup.9, 8.times.10.sup.9,
9.times.10.sup.9, 1.times.10.sup.10, 1.9.times.10.sup.10,
2.times.10.sup.10, 3.times.10.sup.10, 3.73.times.10.sup.10,
4.times.10.sup.10, 5.times.10.sup.10, 6.times.10.sup.10
7.times.10.sup.10, 8.times.10.sup.10, 9.times.10.sup.10,
1.times.10.sup.11, 2.times.10.sup.11, 2.5.times.10.sup.11,
3.times.10.sup.11, 4.times.10.sup.11, 5.times.10.sup.11,
6.times.10.sup.11, 7.times.10.sup.11, 8.times.10.sup.11,
9.times.10.sup.11, 1.times.10.sup.12, 2.times.10.sup.12,
3.times.10.sup.12, 4.times.10.sup.12, 5.times.10.sup.12,
6.times.10.sup.12, 7.times.10.sup.12, 8.times.10.sup.12,
9.times.10.sup.12, 1.times.10.sup.13, 2.times.10.sup.13,
3.times.10.sup.13, 4.times.10.sup.13, 5.times.10.sup.13,
6.times.10.sup.13, 7.times.10.sup.13, 8.times.10.sup.13,
9.times.10.sup.13, 1.times.10.sup.14, 2.times.10.sup.14,
3.times.10.sup.14, 4.times.10.sup.14, 5.times.10.sup.14,
6.times.10.sup.14, 7.times.10.sup.14, 8.times.10.sup.14,
9.times.10.sup.14, 1.times.10.sup.15, 2.times.10.sup.15,
3.times.10.sup.15, 4.times.10.sup.15, 5.times.10.sup.15,
6.times.10.sup.15, 7.times.10.sup.15, 8.times.10.sup.15,
9.times.10.sup.15, or 1.times.10.sup.16 VG. As a non-limiting
example, the total dose is 1.times.10.sup.13 VG. As another
non-limiting example, the total dose is 2.1.times.10.sup.12 VG.
[0415] In one embodiment, about 10.sup.5 to 10.sup.6 viral genome
(unit) may be administered per dose.
[0416] In one embodiment, delivery of the compositions in
accordance with the present disclosure to cells may comprise a
total concentration between about 1.times.10.sup.6 VG/mL and about
1.times.10.sup.16 VG/mL. In some embodiments, delivery may comprise
a composition concentration of about 1.times.10.sup.6,
2.times.10.sup.6, 3.times.10.sup.6, 4.times.10.sup.6,
5.times.10.sup.6, 6.times.10.sup.6, 7.times.10.sup.6,
8.times.10.sup.6, 9.times.10.sup.6, 1.times.10.sup.7,
2.times.10.sup.7, 3.times.10.sup.7, 4.times.10.sup.7,
5.times.10.sup.7, 6.times.10.sup.7, 7.times.10.sup.7,
8.times.10.sup.7, 9.times.10.sup.7, 1.times.10.sup.8,
2.times.10.sup.8, 3.times.10.sup.8, 4.times.10.sup.8,
5.times.10.sup.8, 6.times.10.sup.8, 7.times.10.sup.8,
8.times.10.sup.8, 9.times.10.sup.8, 1.times.10.sup.9,
2.times.10.sup.9, 3.times.10.sup.9, 4.times.10.sup.9,
5.times.10.sup.9, 6.times.10.sup.9, 7.times.10.sup.9,
8.times.10.sup.9, 9.times.10.sup.9, 1.times.10.sup.10,
2.times.10.sup.10, 3.times.10.sup.10, 4.times.10.sup.10,
5.times.10.sup.10, 6.times.10.sup.10, 7.times.10.sup.10,
8.times.10.sup.10, 9.times.10.sup.10, 1.times.10.sup.11,
2.times.10.sup.11, 3.times.10.sup.11, 4.times.10.sup.11,
5.times.10.sup.11, 6.times.10.sup.11, 7.times.10.sup.11,
8.times.10.sup.11, 9.times.10.sup.11, 1.times.10.sup.12,
1.1.times.10.sup.12, 1.2.times.10.sup.12, 1.3.times.10.sup.12,
1.4.times.10.sup.12, 1.5.times.10.sup.12, 1.6.times.10.sup.12,
1.7.times.10.sup.12, 1.8.times.10.sup.12, 1.9.times.10.sup.12,
2.times.10.sup.12, 2.1.times.10.sup.12, 2.2.times.10.sup.12,
2.3.times.10.sup.12, 2.4.times.10.sup.12, 2.5.times.10.sup.12,
2.6.times.10.sup.12, 2.7.times.10.sup.12, 2.8.times.10.sup.12,
2.9.times.10.sup.12, 3.times.10.sup.12, 3.1.times.10.sup.12,
3.2.times.10.sup.12, 3.3.times.10.sup.12, 3.4.times.10.sup.12,
3.5.times.10.sup.12, 3.6.times.10.sup.12, 3.7.times.10.sup.12,
3.8.times.10.sup.12, 3.9.times.10.sup.12, 4.times.10.sup.12,
4.1.times.10.sup.12, 4.2.times.10.sup.12, 4.3.times.10.sup.12,
4.4.times.10.sup.12, 4.5.times.10.sup.12, 4.6.times.10.sup.12,
4.7.times.10.sup.12, 4.8.times.10.sup.12, 4.9.times.10.sup.12,
5.times.10.sup.12, 6.times.10.sup.12, 7.times.10.sup.12,
8.times.10.sup.12, 9.times.10.sup.12, 1.times.10.sup.13,
2.times.10.sup.13, 3.times.10.sup.13, 4.times.10.sup.13,
5.times.10.sup.13, 6.times.10.sup.13, 6.7.times.10.sup.13,
7.times.10.sup.13, 8.times.10.sup.13, 9.times.10.sup.13,
1.times.10.sup.14, 2.times.10.sup.14, 3.times.10.sup.14,
4.times.10.sup.14, 5.times.10.sup.14, 6.times.10.sup.14,
7.times.10.sup.14, 8.times.10.sup.14, 9.times.10.sup.14,
1.times.10.sup.15, 2.times.10.sup.15, 3.times.10.sup.15,
4.times.10.sup.15, 5.times.10.sup.15, 6.times.10.sup.15,
7.times.10.sup.15, 8.times.10.sup.15, 9.times.10.sup.15, or
1.times.10.sup.16 VG/mL. In one embodiment, the concentration of
the AAV particle in the composition is 1.times.10.sup.13 VG/mL. In
one embodiment, the concentration of the AAV particle in the
composition is 3.times.10.sup.12 VG/mL. In one embodiment, the
concentration of the AAV particle in the composition is
1.1.times.10.sup.12 VG/mL. In one embodiment, the concentration of
the AAV particle in the composition is 3.7.times.10.sup.12 VG/mL.
In one embodiment, the concentration of the AAV particle in the
composition is 8.times.10.sup.11 VG/mL. In one embodiment, the
concentration of the AAV particle in the composition is
2.6.times.10.sup.12 VG/mL. In one embodiment, the concentration of
the AAV particle in the composition is 4.9.times.10.sup.12 VG/mL.
In one embodiment, the concentration of the AAV particle in the
composition is 0.8.times.10.sup.12 VG/mL. In one embodiment, the
concentration of the AAV particle in the composition is
0.83.times.10.sup.12 VG/mL. In one embodiment, the concentration of
the AAV particle in the composition is the maximum final dose which
can be contained in a vial.
[0417] In one embodiment, delivery of AAV particles to cells of the
central nervous system (e.g., parenchyma) may comprise a
composition concentration between about 1.times.10.sup.6 VG/mL and
about 1.times.10.sup.16 VG/mL. In some embodiments, delivery may
comprise a composition concentration of about 1.times.10.sup.6,
2.times.10.sup.6, 3.times.10.sup.6, 4.times.10.sup.6,
5.times.10.sup.6, 6.times.10.sup.6, 7.times.10.sup.6,
8.times.10.sup.6, 9.times.10.sup.6, 1.times.10.sup.7,
2.times.10.sup.7, 3.times.10.sup.7, 4.times.10.sup.7,
5.times.10.sup.7, 6.times.10.sup.7, 7.times.10.sup.7,
8.times.10.sup.7, 9.times.10.sup.7, 1.times.10.sup.8,
2.times.10.sup.8, 3.times.10.sup.8, 4.times.10.sup.8,
5.times.10.sup.8, 6.times.10.sup.8, 7.times.10.sup.8,
8.times.10.sup.8, 9.times.10.sup.8, 1.times.10.sup.9,
2.times.10.sup.9, 3.times.10.sup.9, 4.times.10.sup.9,
5.times.10.sup.9, 6.times.10.sup.9, 7.times.10.sup.9,
8.times.10.sup.9, 9.times.10.sup.9, 1.times.10.sup.10,
2.times.10.sup.10, 3.times.10.sup.10, 4.times.10.sup.10,
5.times.10.sup.10, 6.times.10.sup.10, 7.times.10.sup.10,
8.times.10.sup.10, 9.times.10.sup.10, 1.times.10.sup.11,
2.times.10.sup.11, 3.times.10.sup.11, 4.times.10.sup.11,
5.times.10.sup.11, 6.times.10.sup.11, 7.times.10.sup.11,
8.times.10.sup.11, 9.times.10.sup.11, 1.times.10.sup.12,
2.times.10.sup.12, 3.times.10.sup.12, 4.times.10.sup.12,
5.times.10.sup.12, 6.times.10.sup.12, 7.times.10.sup.12,
8.times.10.sup.12, 9.times.10.sup.12, 1.times.10.sup.13,
2.times.10.sup.13, 3.times.10.sup.13, 4.times.10.sup.13,
5.times.10.sup.13, 6.times.10.sup.13, 7.times.10.sup.13,
8.times.10.sup.13, 9.times.10.sup.13, 1.times.10.sup.14,
2.times.10.sup.14, 3.times.10.sup.14 4.times.10.sup.14,
5.times.10.sup.14, 6.times.10.sup.14, 7.times.10.sup.14,
8.times.10.sup.14, 9.times.10.sup.14, 1.times.10.sup.15,
2.times.10.sup.15, 3.times.10.sup.15, 4.times.10.sup.15,
5.times.10.sup.15, 6.times.10.sup.15, 7.times.10.sup.15,
8.times.10.sup.15, 9.times.10.sup.15, or 1.times.10.sup.16 VG/mL.
In one embodiment, the delivery comprises a composition
concentration of 1.times.10.sup.13 VG/mL. In one embodiment, the
delivery comprises a composition concentration of
2.1.times.10.sup.12 VG/mL.
Regimen
[0418] The desired dosage of the AAV particles of the present
disclosure may be delivered only once, three times a day, two times
a day, once a day, every other day, every third day, every week,
every two weeks, every three weeks, or every four weeks. In certain
embodiments, the desired dosage may be delivered using multiple
administrations (e.g., two, three, four, five, six, seven, eight,
nine, ten, eleven, twelve, thirteen, fourteen, or more
administrations). When multiple administrations are employed, split
dosing regimens such as those described herein may be used. As used
herein, a "split dose" is the division of "single unit dose" or
total daily dose into two or more doses, e.g., two or more
administrations of the "single unit dose". As used herein, a
"single unit dose" is a dose of any therapeutic administered in one
dose/at one time/single route/single point of contact, i.e., single
administration event.
[0419] The desired dosage of the AAV particles of the present
disclosure may be administered as a "pulse dose" or as a
"continuous flow". As used herein, a "pulse dose" is a series of
single unit doses of any therapeutic administered with a set
frequency over a period of time. As used herein, a "continuous
flow" is a dose of therapeutic administered continuously for a
period of time in a single route/single point of contact, i.e.,
continuous administration event. A total daily dose, an amount
given or prescribed in 24 hour period, may be administered by any
of these methods, or as a combination of these methods, or by any
other methods suitable for a pharmaceutical administration.
[0420] In one embodiment, delivery of the AAV particles of the
present disclosure to a subject provides regulating activity of
AADC in a subject. The regulating activity may be an increase in
the production of AADC in a subject. The regulating activity can be
for at least 1 month, 2 months, 3 months, 4 months, 5 months, 6
months, 7 months, 8 months, 9 months, 10 months, 11 months, 1 year,
13 months, 14 months, 15 months, 16 months, 17 months, 18 months,
19 months, 20 months, 20 months, 21 months, 22 months, 23 months, 2
years, 3 years, 4 years, 5 years, 6 years, 7 years, 8 years, 9
years, 10 years or more than 10 years.
[0421] In some embodiments, the AAV particle of the present
disclosure may be administered to a subject using a single dose,
one-time treatment. The dose of the one-time treatment may be
administered by any methods known in the art and/or described
herein. As used herein, a "one-time treatment" refers to a
composition which is only administered one time. If needed, a
booster dose may be administered to the subject to ensure the
appropriate efficacy is reached. A booster may be administered 1
month, 2 months, 3 months, 4 months, 5 months, 6 months, 7 months,
8 months, 9 months, 10 months, 11 months, 12 months, 1 year, 13
months, 14 months, 15 months, 16 months, 17 months, 18 months, 19
months, 20 months, 21 months, 22 months, 23 months, 24 months, 2
years, 3 years, 4 years, 5 years, 6 years, 7 years, 8 years, 9
years, 10 years, or more than 10 years after the one-time
treatment.
Delivery Methods
[0422] In one embodiment, the AAV particles or pharmaceutical
compositions of the present disclosure may be administered or
delivered using the methods for treatment of disease described in
U.S. Pat. No. 8,999,948, or International Publication No.
WO2014178863, the contents of which are herein incorporated by
reference in their entirety.
[0423] In one embodiment, the AAV particles or pharmaceutical
compositions of the present disclosure may be administered or
delivered using the methods for delivering gene therapy in
Alzheimer's Disease or other neurodegenerative conditions as
described in US Application No. 20150126590, the contents of which
are herein incorporated by reference in their entirety.
[0424] In one embodiment, the AAV particles or pharmaceutical
compositions of the present disclosure may be administered or
delivered using the methods for delivery of a CNS gene therapy as
described in U.S. Pat. Nos. 6,436,708, and 8,946,152, and
International Publication No. WO2015168666, the contents of which
are herein incorporated by reference in their entirety.
[0425] In one embodiment, the AAV particle comprising an AADC
polynucleotide may be administered or delivered using the methods
for the delivery of AAV virions described in European Patent
Application No. EP1857552, the contents of which are herein
incorporated by reference in its entirety.
[0426] In one embodiment, the AAV particle or pharmaceutical
compositions of the present disclosure may be administered or
delivered using the methods for delivering proteins using AAV
particles described in European Patent Application No. EP2678433,
the contents of which are herein incorporated by reference in their
entirety.
[0427] In one embodiment, the AAV particle comprising an AADC
polynucleotide may be administered or delivered using the methods
for delivering DNA molecules using AAV particles described in U.S.
Pat. No. 5,858,351, the contents of which are herein incorporated
by reference in its entirety.
[0428] In one embodiment, the AAV particle or pharmaceutical
compositions of the present disclosure may be administered or
delivered using the methods for delivering DNA to the bloodstream
described in U.S. Pat. No. 6,211,163, the contents of which are
herein incorporated by reference in their entirety.
[0429] In one embodiment, the AAV particle or pharmaceutical
compositions of the present disclosure may be administered or
delivered using the methods for delivering a payload to the central
nervous system described in U.S. Pat. No. 7,588,757, the contents
of which are herein incorporated by reference in their
entirety.
[0430] In one embodiment, the AAV particle may be administered or
delivered using the methods for delivering AAV virions described in
U.S. Pat. No. 6,325,998, the contents of which are herein
incorporated by reference in its entirety.
[0431] In one embodiment, the AAV particle or pharmaceutical
compositions of the present disclosure may be administered or
delivered using the methods for delivering a payload described in
U.S. Pat. No. 8,283,151, the contents of which are herein
incorporated by reference in their entirety.
[0432] In one embodiment, the AAV particle or pharmaceutical
compositions of the present disclosure may be administered or
delivered using the methods for delivering a payload using a
glutamic acid decarboxylase (GAD) delivery vector described in
International Patent Publication No. WO2001089583, the contents of
which are herein incorporated by reference in their entirety.
[0433] In one embodiment, the AAV particle or pharmaceutical
compositions of the present disclosure may be administered or
delivered using the methods for delivering a payload to neural
cells described in International Patent Publication No.
WO2012057363, the contents of which are herein incorporated by
reference in their entirety.
Delivery to Cells
[0434] The present disclosure provides a method of delivering to a
cell or tissue any of the above-described AAV particles, comprising
contacting the cell or tissue with said AAV particle or contacting
the cell or tissue with a formulation comprising said AAV particle,
or contacting the cell or tissue with any of the described
compositions, including pharmaceutical compositions. The method of
delivering the AAV particle to a cell or tissue can be accomplished
in vitro, ex vivo, or in vivo.
Delivery to Subjects
[0435] The present disclosure additionally provides a method of
delivering to a subject, including a mammalian subject, any of the
above-described AAV particles comprising administering to the
subject said AAV particle, or administering to the subject a
formulation comprising said AAV particle, or administering to the
subject any of the described compositions, including pharmaceutical
compositions.
Combinations
[0436] The AAV particles may be used in combination with one or
more other therapeutic, prophylactic, research or diagnostic
agents. By "in combination with," it is not intended to imply that
the agents must be administered at the same time and/or formulated
for delivery together, although these methods of delivery are
within the scope of the present disclosure. Compositions can be
administered concurrently with, prior to, or subsequent to, one or
more other desired therapeutics or medical procedures. In general,
each agent will be administered at a dose and/or on a time schedule
determined for that agent. In some embodiments, the present
disclosure encompasses the delivery of pharmaceutical,
prophylactic, research, or diagnostic compositions in combination
with agents that may improve their bioavailability, reduce and/or
modify their metabolism, inhibit their excretion, and/or modify
their distribution within the body.
[0437] In one embodiment, the AAV particles described herein may be
administered to a subject who is also undergoing levodopa therapy.
As a non-limiting example, the subject may have a positive response
to levodopa therapy and at least one symptom of PD is reduced. As
another non-limiting example, the subject may have a response to
levodopa therapy where the symptoms of PD experienced by the
subject are stable. As yet another non-limiting example, the
subject may have a negative response to levodopa therapy where the
symptoms of PD experienced by the subject are increasing.
[0438] In one embodiment, the dose of levodopa administered to the
subject prior to the AAV articles is 1, 2, 3, 4, 5, 6, 7, 8, 9, 10,
11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25 or more
than 25 mg/kg. As a non-limiting example, the dose is 3 mg/kg. As
another non-limiting example, the dose is 10 mg/kg. As yet another
non-limiting example, the dose is 20 mg/kg. The subject's response
(e.g., behavioral response) to levodopa may be assessed prior to
administration of the AAV particles. Additionally, the subject may
be administered levodopa again after the administration of the AADC
polynucleotides (e.g., 1 week, 2, weeks, 3 weeks, 1 month, 2
months, 3 months, 4 months, 5 months, 6 months, 7 months, 8 months,
9 months, 10 months, 11 months, 1 year or more than 1 year after
the administration of AAV particles). The behavioral response can
be re-assessed and compared to the initial response to determine
the effects of the AAV particles. The subject may have 1%, 2%, 3%,
4%, 5%, 6%, 7%, 8%, 9%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%,
50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 99% or 100%
behavioral improvement.
[0439] In one embodiment, Levodopa may be administered multiple
times after the administration of the AAV particles. Levodopa may
be administered on a repeating schedule (e.g., every 5 days,
weekly, every 10 days, every 15 days, every 30 days, monthly,
bimonthly, every 3 months, every 4 months, every 5 months, every 6
months, every 7 months, every 8 months, every 9 months, every 10
months, every 11 months or yearly) or as symptoms arise. As a
non-limiting example, 3 years post administration of AADC
polynucleotides a subject may have 1-10%, 5-15%, 10-20%, 15-30%,
20-40%, 25-50%, 30-50%, 40-50%, 40-60%, 50-70%, 50-80%, 60-70%,
60-75%, 60-80%, 60-90%, 70-80%, 70-90%, 75-90%, 80-90%, 90-100% of
the striatal neurons within the infused region of the putamen to be
AADC-immunoreactive. As a non-limiting example, 6 years post
administration of AAV particles a subject may have 1-10%, 5-15%,
10-20%, 15-30%, 20-40%, 25-50%, 30-50%, 40-50%, 40-60%, 50-70%,
50-80%, 60-70%, 60-75%, 60-80%, 60-90%, 70-80%, 70-90%, 75-90%,
80-90%, 90-100% of the striatal neurons within the infused region
of the putamen to be AADC-immunoreactive. As a non-limiting
example, 9 years post administration of AADC polynucleotides a
subject may have 1-10%, 5-15%, 10-20%, 15-30%, 20-40%, 25-50%,
30-50%, 40-50%, 40-60%, 50-70%, 50-80%, 60-70%, 60-75%, 60-80%,
60-90%, 70-80%, 70-90%, 75-90%, 80-90%, 90-100% of the striatal
neurons within the infused region of the putamen to be
AADC-immunoreactive.
[0440] In one embodiment, a subject who may be administered the AAV
particles described herein have a documented response to levodopa
therapy but have medically refractory fluctuations and are
considered good surgical candidates. The determination if a subject
is a good surgical candidate may be made by the physician treating
the subject for PD or the physician administering the AAV particles
who takes into consideration the overall risk to benefit ratio for
the patient for the surgical intervention required for delivery of
the AAV particles.
Measurement of Expression
[0441] Expression of payloads from viral genomes may be determined
using various methods known in the art such as, but not limited to
immunochemistry (e.g., IHC), in situ hybridization (ISH),
enzyme-linked immunosorbent assay (ELISA), affinity ELISA, ELISPOT,
flow cytometry, immunocytology, surface plasmon resonance analysis,
kinetic exclusion assay, liquid chromatography-mass spectrometry
(LCMS), high-performance liquid chromatography (HPLC), BCA assay,
immunoelectrophoresis, Western blot, SDS-PAGE, protein
immunoprecipitation, and/or PCR.
[0442] The pharmaceutical compositions of AAV particles described
herein may be characterized by one or more of bioavailability,
therapeutic window and/or volume of distribution.
Bioavailability
[0443] The AAV particles, when formulated into a composition with a
delivery agent as described herein, can exhibit an increase in
bioavailability as compared to a composition lacking a delivery
agent as described herein. As used herein, the term
"bioavailability" refers to the systemic availability of a given
amount of AAV particle or expressed payload administered to a
mammal. Bioavailability can be assessed by measuring the area under
the curve (AUC) or the maximum serum or plasma concentration
(C.sub.max) of the composition following. AUC is a determination of
the area under the curve plotting the serum or plasma concentration
of a compound (e.g., AAV particles or expressed payloads) along the
ordinate (Y-axis) against time along the abscissa (X-axis).
Generally, the AUC for a particular compound can be calculated
using methods known to those of ordinary skill in the art and as
described in G. S. Banker, Modern Pharmaceutics, Drugs and the
Pharmaceutical Sciences, v. 72, Marcel Dekker, New York, Inc.,
1996, the contents of which are herein incorporated by reference in
its entirety.
[0444] The C.sub.max value is the maximum concentration of the AAV
particle or expressed payload achieved in the serum or plasma of a
mammal following administration of the AAV particle to the mammal.
The C.sub.max value of can be measured using methods known to those
of ordinary skill in the art. The phrases "increasing
bioavailability" or "improving the pharmacokinetics," as used
herein mean that the systemic availability of a first AAV particle
or expressed payload, measured as AUC, C.sub.max, or C.sub.min in a
mammal is greater, when co-administered with a delivery agent as
described herein, than when such co-administration does not take
place. In some embodiments, the bioavailability can increase by at
least about 2%, at least about 5%, at least about 10%, at least
about 15%, at least about 20%, at least about 25%, at least about
30%, at least about 35%, at least about 40%, at least about 45%, at
least about 50%, at least about 55%, at least about 60%, at least
about 65%, at least about 70%, at least about 75%, at least about
80%, at least about 85%, at least about 90%, at least about 95%, or
about 100%.
Therapeutic Window
[0445] As used herein "therapeutic window" refers to the range of
plasma concentrations, or the range of levels of therapeutically
active substance at the site of action, with a high probability of
eliciting a therapeutic effect. In some embodiments, the
therapeutic window of the AAV particle as described herein can
increase by at least about 2%, at least about 5%, at least about
10%, at least about 15%, at least about 20%, at least about 25%, at
least about 30%, at least about 35%, at least about 40%, at least
about 45%, at least about 50%, at least about 55%, at least about
60%, at least about 65%, at least about 70%, at least about 75%, at
least about 80%, at least about 85%, at least about 90%, at least
about 95%, or about 100%.
Volume of Distribution
[0446] As used herein, the term "volume of distribution" refers to
the fluid volume that would be required to contain the total amount
of the drug in the body at the same concentration as in the blood
or plasma: V.sub.dist equals the amount of drug in the
body/concentration of drug in blood or plasma. For example, for a
10 mg dose and a plasma concentration of 10 mg/L, the volume of
distribution would be 1 liter. The volume of distribution reflects
the extent to which the drug is present in the extravascular
tissue. A large volume of distribution reflects the tendency of a
compound to bind to the tissue components compared with plasma
protein binding. In a clinical setting, V.sub.dist can be used to
determine a loading dose to achieve a steady state concentration.
In some embodiments, the volume of distribution of the AAV
particles as described herein can decrease at least about 2%, at
least about 5%, at least about 10%, at least about 15%, at least
about 20%, at least about 25%, at least about 30%, at least about
35%, at least about 40%, at least about 45%, at least about 50%, at
least about 55%, at least about 60%, at least about 65%, at least
about 70%.
Biological Effect
[0447] In one embodiment, the biological effect of the AAV
particles delivered to the animals may be categorized by analyzing
the payload expression in the animals. The payload expression may
be determined from analyzing a biological sample collected from a
mammal administered the AAV particles of the present disclosure.
For example, a protein expression of 50-200 pg/ml for the protein
encoded by the AAV particles delivered to the mammal may be seen as
a therapeutically effective amount of protein in the mammal.
IV. Methods and Uses of the Compositions
CNS Diseases
[0448] The polynucleotides of the present disclosure may be used in
the treatment, prophylaxis or amelioration of any disease or
disorder characterized by aberrant or undesired target expression.
In one embodiment, the disclosure relates to AAV particles for use
in the treatment of Parkinson's disease.
[0449] In some embodiments, the AAV particles may be used in the
treatment, prophylaxis or amelioration of any disease or disorder
characterized by aberrant or undesired target expression wherein
the payload, i.e. AADC, is swapped for an alternate payload.
[0450] The present disclosure provides a method for treating a
disease, disorder and/or condition in a mammalian subject,
including a human subject, comprising administering to the subject
AAV particles described herein.
[0451] In one embodiment, the disease, disorder and/or condition is
a neurological disease, disorder and/or condition. The CNS diseases
may be diseases that affect any component of the brain (including
the cerebral hemispheres, diencephalon, brain stem, and cerebellum)
or the spinal cord.
[0452] In some embodiments, viral particles of the present
disclosure, through delivery of a functional payload that is a
therapeutic product that can modulate the level or function of a
gene product in the CNS, may be used to treat a neurodegenerative
diseases and/or diseases or disorders that are characteristic with
neurodegeneration, neuromuscular diseases, lysosomal diseases,
trauma, bone marrow injuries, pain (including neuropathic pain),
cancers of the nervous system, demyelinating diseases, autoimmune
diseases of the nervous system, neurotoxic syndromes, sleeping
disorders, genetic brain disorders and developmental CNS disorders.
A functional payload may alleviate or reduce symptoms that result
from abnormal level and/or function of a gene product (e.g., an
absence or defect in a protein) in a subject in need thereof or
that otherwise confers a benefit to a CNS disorder in a subject in
need thereof.
[0453] As non-limiting examples, therapeutic products delivered by
AAV particles of the present disclosure may include, but are not
limited to, growth and trophic factors, cytokines, hormones,
neurotransmitters, enzymes, anti-apoptotic factors, angiogenic
factors, and any protein known to be mutated in pathological
disorders such as the "survival of motor neuron" protein (SMN);
antisense RNA or RNAi targeting messenger RNAs coding for proteins
having a therapeutic interest in any of CNS diseases discussed
herein; or microRNAs that function in gene silencing and
post-transcriptionally regulation of gene expression in the CNS
(e.g., brain specific Mir-128a, See Adlakha and Saini, Molecular
cancer, 2014, 13:33). For example, an RNAi targeting the superoxide
dismutase enzyme may be packaged by viral particles of the present
disclosure, for the treatment of ALS.
[0454] The growth and trophic factors may include, but are not
limited to brain-derived growth factor (BDNF), epidermal growth
factor (EGF), basic Fibroblast growth factor (bFGF), Ciliary
neurotrophic factor (CNTF), corticotropin-releasing factor (CRF),
Glial cell line derived growth factor (GDNF), Insulin-like growth
factor-1 (IGF-1), nerve growth factor (NGF), neurotrophin-3 (NT-3),
neurotrophin-4 (NT-4), and vascular endothelial growth factor
(VEGF). Cytokines may include interleukin-10 (IL-10),
interleukin-6, Interleukin-8, chemokine CXCL12 (SDF-1), TGF-beta,
and Growth and differentiation factor (GDF-1/10).
[0455] In some embodiments, the neurological disorders may be
neurodegenerative disorders including, but not limited to,
Alzheimer's Diseases (AD); Amyotrophic lateral sclerosis (ALS);
Creutzfeldt-Jakob Disease (CJD); Huntingtin's disease (HD);
Friedreich's ataxia (FA); Parkinson Disease (PD); Multiple System
Atrophy (MSA); Spinal Muscular Atrophy (SMA), Multiple Sclerosis
(MS); Primary progressive aphasia; Progressive supranuclear palsy
(PSP); Dementia; Brain Cancer, Degenerative Nerve Diseases,
Encephalitis, Epilepsy, Genetic Brain Disorders that cause
neurodegeneration, Retinitis pigmentosa (RP), Head and Brain
Malformations, Hydrocephalus, Stroke, Prion disease, Infantile
neuronal ceroid lipofuscinosis (INCL) (a neurodegenerative disease
of children caused by a deficiency in the lysosomal enzyme
palmitoyl protein thioesterase-1 (PPT1)), and others.
[0456] In some embodiments, viral particles of the present
disclosure may be used to treat diseases that are associated with
impairments of the growth and development of the CNS, i.e.,
neurodevelopmental disorders. In some aspects, such
neurodevelopmental disorders may be caused by genetic mutations,
including but not limited to, Fragile X syndrome (caused by
mutations in FMR1 gene), Down syndrome (caused by trisomy of
chromosome 21), Rett syndrome, Williams syndrome, Angelman
syndrome, Smith-Magenis syndrome, ATR-X syndrome, Barth syndrome,
Immune dysfunction and/or infectious diseases during infancy such
as Sydenham's chorea, Schizophrenia Congenital toxoplasmosis,
Congenital rubella syndrome, Metabolic disorders such as diabetes
mellitus and phenylketonuria; nutritional defects and/or brain
trauma, Autism and autism spectrum.
[0457] In some embodiments, viral particles of the present
disclosure, may be used to treat a tumor in the CNS, including but
not limited to, acoustic neuroma, Astrocytoma (Grades I, II, III
and IV), Chordoma, CNS Lymphoma, Craniopharyngioma, Gliomas (e.g.,
brain stem glioma, ependymoma, optical nerve glioma,
subependymoma), Medulloblastoma, Meningioma, Metastatic brain
tumors, Oligodendroglioma, Pituitary Tumors, Primitive
neuroectodermal (PNET), and Schwannoma.
[0458] In some embodiments, the neurological disorders may be
functional neurological disorders with motor and/or sensory
symptoms which have neurological origin in the CNS. As non-limiting
examples, functional neurological disorders may be chronic pain,
seizures, speech problems, involuntary movements, and sleep
disturbances.
[0459] In some embodiments, the neurological disorders may be white
matter disorders (a group of diseases that affects nerve fibers in
the CNS) including but not limited to, Pelizaeus-Merzbacher
disease, Hypomyelination with atrophy of basal ganglia and
cerebellum, Aicardi-Goutieres syndrome, Megalencephalic
leukoencephalopathy with subcortical cysts, Congenital muscular
dystrophies, Myotonic dystrophy, Wilson disease, Lowe syndrome,
Sjogren-Larsson syndrome, PIBD or Tay syndrome, Cockayne's disease,
erebrotendinous xanthomatosis, Zellweger syndrome, Neonatal
adrenoleukodystrophy, Infantile Refsum disease, Zellweger-like
syndrome, Pseudo-Zellweger syndrome, Pseudo-neonatal
adrenoleukodystrophy, Bifunctional protein deficiency, X-linked
adrenoleukodystrophy and adrenomyeloneuropathy and Refsum
disease.
[0460] In some embodiments, the neurological disorders may be
lysosomal storage disorders (LSDs) caused by the inability of cells
in the CNS to break down metabolic end products, including but not
limited to Niemann-Pick disease (a LSD resulting from inherited
deficiency in acid sphingomyelinase (ASM); Metachromatic
leukodystrophy (MLD) (a LSD characterized by accumulation of
sulfatides in glial cells and neurons, the result of an inherited
deficiency of arylsulfatase A (ARSA)); Globoid-cell leukodystrophy
(GLD) (a LSD caused by mutations in galactosylceramidase); Fabry
disease (a LSD caused by mutations in the alpha-galactosidase A
(GLA) gene); Gaucher disease (caused by mutations in the
beta-glucocerebrosidase (GBA) gene); GM1/GM2 gangliosidosis;
Mucopolysaccharidoses disorder; Pompe disease; and Neuronal ceroid
lipofuscinosis.
[0461] In another embodiment, the neurological disease, disorder
and/or condition is Friedreich's Ataxia. In one embodiment, the AAV
particle used to treat Friedreich's Ataxia comprises a nucleic acid
sequence such as, but not limited to, SEQ ID NO: 979 or a fragment
or variant thereof, wherein the payload is replaced by Frataxin or
any other payload known in the art for treating Friedreich's
Ataxia.
[0462] In another embodiment, the neurological disease, disorder
and/or condition is Amyotrophic lateral sclerosis (ALS). In one
embodiment the AAV particle used to treat ALS comprises a nucleic
acid sequence such as, but not limited to, SEQ ID NO: 979 or a
fragment or variant thereof, wherein the payload is replaced by
replaced by an shRNA, miRNA, siRNA, RNAi for SOD1 or any other
payload known in the art for treating ALS.
[0463] In another embodiment, the neurological disease, disorder
and/or condition is Huntington's disease. In one embodiment the AAV
particle used to treat Huntington's disease comprises a nucleic
acid sequence such as, but not limited to, SEQ ID NO: 979 or a
fragment or variant thereof, wherein the payload is replaced by
replaced by an shRNA, miRNA, siRNA, RNAi for Htt or any other
payload known in the art for treating Huntington's disease.
[0464] In another embodiment, the neurological disease, disorder or
condition is spinal muscular atrophy (SMA). In another embodiment,
the neurological disease, disorder and/or condition is Friedreich's
Ataxia. In one embodiment the AAV particle used to treat SMN
comprises a nucleic acid sequence such as, but not limited to, SEQ
ID NO: 979 or a fragment or variant thereof, wherein the payload is
replaced by Frataxin or any other payload known in the art for
treating SMA.
Parkinson's Disease
[0465] In one embodiment, the neurological disease, disorder and/or
condition is Parkinson's disease. In one embodiment the AAV
particle used to treat Parkinson's disease comprises a payload such
as, but not limited to, SEQ ID NO: 979 or a fragment or variant
thereof.
[0466] In one embodiment, the subject is a human patient who has a
minimum motor score of about 30 to a maximum score of about 100,
about 10 to a maximum score of about 100, about 20 to a maximum
score of about 100 in the Unified Parkinson's Disease Rating
Scale.
[0467] In one embodiment, the subject has been diagnosed with
Parkinson's disease within the past 5 years prior to treatment with
the compositions described herein. As a non-limiting example, the
subject may have been diagnosed with Parkinson's disease within a
week, 2 weeks, 3 weeks, 4 weeks, 5 weeks, 6 weeks, 7 weeks, 8
weeks, 1 month, 2 months, 3 months, 4 months, 5 months, 6
[0468] months, 7 months, 8 months, 9 months, 10 months, 11 months,
12 months, 13 months, 14 months, 15 months, 16 months, 17 months,
18 months, 1 year, 2 years, 3 years, 4 years or less than 5 years
prior to treatment with the compositions described herein.
[0469] In one embodiment, the subject has been diagnosed with
Parkinson's disease between 5 and 10 years prior to treatment with
the compositions described herein. As a non-limiting example, the
subject may have been diagnosed with Parkinson's disease 5, 5.5.,
6, 6.5, 7, 7.5, 8, 8.5, 9, 9.5 or 10 years prior to treatment with
the compositions described herein.
[0470] In one embodiment, the subject has been diagnosed with
Parkinson's disease more than 10 years prior to treatment with the
compositions described herein. As a non-limiting example, the
subject may have been diagnosed with Parkinson's disease 10.5, 11,
11.5, 12, 12.5, 13, 13.5, 14, 14.5, 15, 15.5, 16, 16.5, 17, 17.5,
18, 18.5, 19, 19.5, 20, 20.5, 21, 21.5, 22, 22.5, 23, 23.5, 24 or
more than 24 years prior to treatment with the compositions
described herein. In one embodiment, a subject is 50-65 years of
age. As a non-limiting example, the subject is 50, 51, 52, 53, 54,
55, 56, 57, 58, 59, 60, 61, 62, 63, 64, or 65 years of age. As a
non-limiting example, the subject is 50 years of age. As a
non-limiting example, the subject is 51 years of age. As a
non-limiting example, the subject is 52 years of age. As a
non-limiting example, the subject is 53 years of age. As a
non-limiting example, the subject is 54 years of age. As a
non-limiting example, the subject is 55 years of age. As a
non-limiting example, the subject is 56 years of age. As a
non-limiting example, the subject is 57 years of age. As a
non-limiting example, the subject is 58 years of age. As a
non-limiting example, the subject is 59 years of age. As a
non-limiting example, the subject is 60 years of age. As a
non-limiting example, the subject is 61 years of age. As a
non-limiting example, the subject is 62 years of age. As a
non-limiting example, the subject is 63 years of age. As a
non-limiting example, the subject is 64 years of age. As a
non-limiting example, the subject is 65 years of age.
[0471] In one embodiment, a subject is 30 to 50 years of age. As a
non-limiting example, the subject is 30 years of age. As a
non-limiting example, the subject is 31 years of age. As a
non-limiting example, the subject is 32 years of age. As a
non-limiting example, the subject is 33 years of age. As a
non-limiting example, the subject is 34 years of age. As a
non-limiting example, the subject is 35 years of age. As a
non-limiting example, the subject is 36 years of age. As a
non-limiting example, the subject is 37 years of age. As a
non-limiting example, the subject is 38 years of age. As a
non-limiting example, the subject is 39 years of age. As a
non-limiting example, the subject is 40 years of age. As a
non-limiting example, the subject is 41 years of age. As a
non-limiting example, the subject is 42 years of age. As a
non-limiting example, the subject is 43 years of age. As a
non-limiting example, the subject is 44 years of age. As a
non-limiting example, the subject is 45 years of age. As a
non-limiting example, the subject is 46 years of age. As a
non-limiting example, the subject is 47 years of age. As a
non-limiting example, the subject is 48 years of age. As a
non-limiting example, the subject is 49 years of age. As a
non-limiting example, the subject is 50 years of age.
[0472] In one embodiment, a subject is 65 to 85 years of age. As a
non-limiting example, the subject is 65 years of age. As a
non-limiting example, the subject is 66 years of age. As a
non-limiting example, the subject is 67 years of age. As a
non-limiting example, the subject is 68 years of age. As a
non-limiting example, the subject is 69 years of age. As a
non-limiting example, the subject is 70 years of age. As a
non-limiting example, the subject is 71 years of age. As a
non-limiting example, the subject is 72 years of age. As a
non-limiting example, the subject is 73 years of age. As a
non-limiting example, the subject is 74 years of age. As a
non-limiting example, the subject is 75 years of age. As a
non-limiting example, the subject is 76 years of age. As a
non-limiting example, the subject is 77 years of age. As a
non-limiting example, the subject is 78 years of age. As a
non-limiting example, the subject is 79 years of age. As a
non-limiting example, the subject is 80 years of age. As a
non-limiting example, the subject is 81 years of age. As a
non-limiting example, the subject is 82 years of age. As a
non-limiting example, the subject is 83 years of age. As a
non-limiting example, the subject is 84 years of age. As a
non-limiting example, the subject is 85 years of age.
[0473] In one embodiment, a subject has seen a change in motor
symptoms such as tremors and movements prior to administration of
the composition described herein. Non-limiting examples of tremors
include, unilateral or bilateral mild tremors, bilateral or midline
moderate tremors or intractable tremors. Non-limiting examples of
movements include mild bradykinesia, moderate bradykinesia, severe
bradykinesia and morning akinesia.
[0474] In one embodiment, a subject may have changes in balance
such as, but not limited to, impaired balance, impaired righting
reflexes, significant balance disorder or falling.
[0475] In one embodiment, a subject may have a reduced quality of
life. As a non-limiting example, the subject may have a moderate
impact on their quality of life such as experiencing some
limitations to activities of daily living. As another non-limiting
example, the subject may have a quality of life which has been
diminished by illness.
[0476] In one embodiment, a subject has seen a change in non-motor
symptoms prior to administration of the composition described
herein. As a non-limiting example, the subject may have mild to
moderate cognitive impairment prior to administration to the
composition described herein. As another non-limiting example, the
subject may have significant cognitive impairment such as dementia
which may also include behavioral disturbances such as
hallucinations.
[0477] In one embodiment, a subject may have a satisfactory
response with limited fluctuations on one or more dopaminergic
medications prior to administration of the compositions described
herein.
[0478] In one embodiment, a subject may have motor fluctuations
causing mild to moderate disability on one or more dopaminergic
medications prior to administration of the compositions described
herein.
[0479] In one embodiment, a subject may have medically refractory
motor fluctuations consisting of "wearing off" and/or
levodopa-induced dyskinesias causing significant disability prior
to administration of the compositions described herein.
[0480] In one embodiment, a subject may have mild symptoms
associated with Parkinson's disease such as, but not limited to, no
cognitive impairment, diagnosed within the past 5 years,
satisfactory response with limited fluctuations on one or more
dopaminergic medications, unilateral or bilateral mild tremors,
little to no impact on the quality of life, and/or no balance
impairment.
[0481] In one embodiment, a subject may have moderate symptoms
associated with Parkinson's disease such as, but not limited to,
mild to moderate cognitive impairment, first signs of impaired
balance and righting reflexes, motor fluctuations causing
mild-moderate disability on one or more dopaminergic medications,
diagnosed within the past 5 to 10 years, bilateral or midline
moderate tremors, moderate bradykinesia and/or subject experiencing
some limitations to activities of daily living.
[0482] In one embodiment, a subject may have advanced symptoms
associated with Parkinson's disease such as, but not limited to,
being diagnosed with Parkinson's more than 10 years, medium
refractory motor fluctuations wearing off and/or levodopa-induced
dyskinesia causing significant disability, intractable tremors,
significant balance disorder and/or falling, significant cognitive
impairment (such as dementia with or without behavioral
disturbances), sever bradykinesia, quality of life markedly
diminished by illness and/or morning akinesia.
[0483] In one embodiment, a subject has been referred to a movement
disorder specialist (MDS) but has not undergone deep brain
stimulation.
[0484] In one embodiment, a subject is using DUOPA.TM. in
combination with the compositions described herein. As a
non-limiting example, the subject may have success with using
DUOPA.TM. alone. As a non-limiting example, the subject may not
have any success or limited success using DUOPA.TM. alone.
[0485] In one embodiment, a subject is one who was a candidate for
surgical intervention including, but not limited to, deep-brain
stimulation. As a non-limiting example, deep-brain stimulation was
suggested due to disabling motor complications despite treatment
with optimal anti-Parkinsonian medication.
[0486] In one embodiment, a subject has an average on-time of
7.5-14 hours based on the subject diary. As a non-limiting example,
the average on-time is 7.5, 7.6, 7.7, 7.8, 7.9, 8, 8.1, 8.2, 8.3,
8.4, 8.5, 8.6, 8.7, 8.8, 8.9, 9, 9.1, 9.2, 9.3, 9.4, 9.5, 9.6, 9.7,
9.8, 9.9, 10, 10.1, 10.2, 10.3, 10.4, 10.5, 10.6, 10.7, 10.8, 10.9,
11, 11.1, 11.2, 11.3, 11.4, 11.5, 11.6, 11.7, 11.8, 11.9, 12, 12.1,
12.2, 12.3, 12.4, 12.5, 12.6, 12.7, 12.8, 12.9, 13, 13.1, 13.2,
13.3, 13.4, 13.5, 13.6, 13.7, 13.8, 13.9, or 14 hours. In one
embodiment, a subject has an average on-time of 10.5 hours based on
the subject diary.
[0487] In one embodiment, a subject experiences about 1.5 hours
more of diary on-time (without troublesome dyskinesia), as compared
to baseline, 6 months after administration of the present
disclosure. As a non-limiting example, a subject experiences about
1.5 hours more of diary on-time 6 months after administration of
the present disclosure at a dose volume of up to 900 uL per putamen
and a total dose of 4.5.times.10.sup.12 vector genome.
[0488] In one embodiment, a subject experiences about 2.2 hours
more of diary on-time (without troublesome dyskinesia), as compared
to baseline, 6 months after administration of the present
disclosure. As a non-limiting example, a subject experiences about
2.2 hours more of diary on-time 6 months after administration of
the present disclosure at a dose volume of up to 900 uL per putamen
and a total dose of 1.5.times.10.sup.12 vector genome.
[0489] In one embodiment, a subject experiences about 4 hours more
of diary on-time (without troublesome dyskinesia), as compared to
baseline, 6 months after administration of the present disclosure.
As a non-limiting example, a subject experiences about 4 hours more
of diary on-time 6 months after administration of the present
disclosure at a dose volume of up to 900 uL per putamen and a total
dose of 1.5.times.10.sup.12 vector genome.
[0490] In one embodiment, a subject experiences about 1.6 hours
more of diary on-time (without troublesome dyskinesia), as compared
to baseline, 12 months after administration of the present
disclosure. As a non-limiting example, a subject experiences about
1.6 hours more of diary on-time 12 months after administration of
the present disclosure at a dose volume of up to 450 uL per putamen
and a total dose of 7.5.times.10.sup.11 vector genome.
[0491] In one embodiment, a subject experiences about 3.3 hours
more of diary on-time (without troublesome dyskinesia), as compared
to baseline, 12 months after administration of the present
disclosure. As a non-limiting example, a subject experiences about
3.3 hours more of diary on-time 12 months after administration of
the present disclosure at a dose volume of up to 900 uL per putamen
and a total dose of 1.5.times.10.sup.12 vector genome.
[0492] In one embodiment, a subject experiences about 4 hours more
of diary on-time (without troublesome dyskinesia), as compared to
baseline, 12 months after administration of the present disclosure.
As a non-limiting example, a subject experiences about 4 hours more
of diary on-time 12 months after administration of the present
disclosure at a dose volume of up to 900 uL per putamen and a total
dose of 1.5.times.10.sup.12 vector genome.
[0493] In one embodiment, a subject experiences about 2.3 hours
more of diary on-time (without troublesome dyskinesia), as compared
to baseline, 24 months after administration of the present
disclosure. As a non-limiting example, a subject experiences about
2.3 hours more of diary on-time 24 months after administration of
the present disclosure at a dose volume of up to 450 uL per putamen
and a total dose of 7.5.times.10.sup.11 vector genome.
[0494] In one embodiment, a subject has an average off-time of 2-7
hours based on the subject diary. As a non-limiting example, the
average off-time is 2, 2.1, 2.2, 2.3, 2.4, 2.5, 2.6, 2.7, 2.8, 2.9,
3, 3.1, 3.2, 3.3, 3.4, 3.5, 3.6, 3.7, 3.8, 3.9, 4, 4.1, 4.2, 4.3,
4.4, 4.5, 4.6, 4.7, 4.8, 4.9, 5, 5.1, 5.2, 5.3, 5.4, 5.5, 5.6, 5.7,
5.8, 5.9, 6, 6.1, 6.2, 6.3, 6.4, 6.5, 6.6, 6.7, 6.8, 6.9, or 7. In
one embodiment, a subject has an average off-time of 4.6 hours
based on the subject diary.
[0495] In one embodiment, a subject experiences about 1.3 hours
less of diary off-time, as compared to baseline, 6 months after
administration of the present disclosure. As a non-limiting
example, a subject experiences about 1.3 hours less of diary
off-time 6 months after administration of the present disclosure at
a dose volume of up to 900 uL per putamen and a total dose of
4.5.times.10.sup.12 vector genome.
[0496] In one embodiment, a subject experiences about 1.1 hours
less of diary off-time, as compared to baseline, 6 months after
administration of the present disclosure. As a non-limiting
example, a subject experiences about 1.1 hours less of diary
off-time 6 months after administration of the present disclosure at
a dose volume of up to 900 uL per putamen and a total dose of
1.5.times.10.sup.12 vector genome.
[0497] In one embodiment, a subject experiences about 0.8 hours
less of diary off-time, as compared to baseline, 6 months after
administration of the present disclosure. As a non-limiting
example, a subject experiences about 0.8 hours less of diary
off-time 6 months after administration of the present disclosure at
a dose volume of up to 450 uL per putamen and a total dose of
7.5.times.10.sup.11 vector genome.
[0498] In one embodiment, a subject experiences about 2.3 hours
less of diary off-time, as compared to baseline, 12 months after
administration of the present disclosure. As a non-limiting
example, a subject experiences about 2.3 hours less of diary
off-time 12 months after administration of the present disclosure
at a dose volume of up to 900 uL per putamen and a total dose of
1.5.times.10.sup.12 vector genome.
[0499] In one embodiment, a subject experiences about 1.4 hours
less of diary off-time, as compared to baseline, 12 months after
administration of the present disclosure. As a non-limiting
example, a subject experiences about 1.4 hours less of diary
off-time 12 months after administration of the present disclosure
at a dose volume of up to 450 uL per putamen and a total dose of
7.5.times.10.sup.11 vector genome.
[0500] In one embodiment, a subject experiences about 1.8 hours
less of diary off-time, as compared to baseline, 24 months after
administration of the present disclosure. As a non-limiting
example, a subject experiences about 1.8 hours less of diary
off-time 24 months after administration of the present disclosure
at a dose volume of up to 450 uL per putamen and a total dose of
7.5.times.10.sup.11 vector genome.
[0501] In one embodiment, a subject experiences 10% less diary
off-time 6 months after administration of the present disclosure.
In one embodiment, a subject experiences 10% less diary off-time 12
months after administration of the present disclosure.
[0502] In one embodiment, a subject experiences 20% less diary
off-time 6 months after administration of the present disclosure.
In one embodiment, a subject experiences 20% less diary off-time 12
months after administration of the present disclosure.
[0503] In one embodiment, a subject experiences 30% less diary
off-time 6 months after administration of the present disclosure.
In one embodiment, a subject experiences 30% less diary off-time 12
months after administration of the present disclosure.
[0504] In one embodiment, a subject experiences 40% less diary
off-time 6 months after administration of the present disclosure.
In one embodiment, a subject experiences 40% less diary off-time 12
months after administration of the present disclosure.
[0505] In one embodiment, a subject experiences 50% less diary
off-time 6 months after administration of the present disclosure.
In one embodiment, a subject experiences 50% less diary off-time 12
months after administration of the present disclosure.
[0506] In one embodiment, a subject experiences 60% less diary
off-time 6 months after administration of the present disclosure.
In one embodiment, a subject experiences 60% less diary off-time 12
months after administration of the present disclosure.
[0507] In one embodiment, a subject experiences 70% less diary
off-time 6 months after administration of the present disclosure.
In one embodiment, a subject experiences 70% less diary off-time 12
months after administration of the present disclosure.
[0508] In one embodiment, a subject experiences 80% less diary
off-time 6 months after administration of the present disclosure.
In one embodiment, a subject experiences 80% less diary off-time 12
months after administration of the present disclosure.
[0509] In one embodiment, a subject experiences 90% less diary
off-time 6 months after administration of the present disclosure.
In one embodiment, a subject experiences 90% less diary off-time 12
months after administration of the present disclosure.
[0510] In one embodiment, a subject experiences 95% less diary
off-time 6 months after administration of the present disclosure.
In one embodiment, a subject experiences 95% less diary off-time 12
months after administration of the present disclosure.
[0511] In one embodiment, a subject's UPDRS-3 (or UPDRS-III)
medication score is evaluated prior to administration of the
present disclosure. As a non-limiting example, the subject's
UPDRS-3 (or UPDRS-III) medication score prior to administration of
the present disclosure is between 1-40, 1-10, 1-15, 1-20, 1-25,
1-30, 1-35, 1-40, 5-10, 5-15, 5-20, 5-25, 5-30, 5-35, 5-40, 10-15,
10-20, 10-25, 10-30, 10-35, 10-40, 15-20, 15-25, 15-30, 15-35,
15-40, 20-25, 20-30, 20-35, 20-40, 25-30, 25-35, 25-40, 30-35,
30-40, or 35-40. As a non-limiting example, the subject's UPDRS-3
(or UPDRS-III) medication score prior to administration of the
present disclosure is 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13,
14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30,
31, 32, 33, 34, 35, 36, 37, 38, 39, or 40. As a non-limiting
example, the subject's UPDRS-3 (or UPDRS-III) medication score
prior to administration of the present disclosure is 10.1, 10.2,
10.3, 10.4, 10.5, 10.6, 10.7, 10.8, 10.9, 11, 11.1, 11.2, 11.3,
11.4, 11.5, 11.6, 11.7, 11.8, 11.9, 12, 12.1, 12.2, 12.3, 12.4,
12.5, 12.6, 12.7, 12.8, 12.9, 13, 13.1, 13.2, 13.3, 13.4, 13.5,
13.6, 13.7, 13.8, 13.9, 14, 14.1, 14.2, 14.3, 14.4, 14.5, 14.6,
14.7, 14.8, 14.9, or 15. As a non-limiting example, the subjects
UPDRS-3 medication score prior to administration of the present
disclosure is 13.5.
[0512] In one embodiment, a subject's UPDRS-3 (or UPDRS-III)
medication score is reduced after administration of the present
disclosure.
[0513] A subject's UPDRS-3 (or UPDRS-III) medication score may be
reduced by a percentage such as, but not limited to, 10%, 15%, 20%,
25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%,
90%, 95% or more than 95%. As a non-limiting example, a subject's
UPDRS-3 score is reduced 10% after administration of the present
disclosure. As a non-limiting example, a subject's UPDRS-3 score is
reduced 20% after administration of the present disclosure. As a
non-limiting example, a subject's UPDRS-3 score is reduced 30%
after administration of the present disclosure. As a non-limiting
example, a subject's UPDRS-3 score is reduced 40% after
administration of the present disclosure. As a non-limiting
example, a subject's UPDRS-3 score is reduced 50% after
administration of the present disclosure. As a non-limiting
example, a subject's UPDRS-3 score is reduced 60% after
administration of the present disclosure. As a non-limiting
example, a subject's UPDRS-3 score is reduced 70% after
administration of the present disclosure. As a non-limiting
example, a subject's UPDRS-3 score is reduced 80% after
administration of the present disclosure. As a non-limiting
example, a subject's UPDRS-3 score is reduced 90% after
administration of the present disclosure.
[0514] A subject's UPDRS-3 (or UPDRS-III) medication score may
change by 0.1, 0.2, 0.3, 0.4, 0.5, 0.6, 0.7, 0.8, 0.9, 1, 1.1, 1.2,
1.3, 1.4, 1.5, 1.6, 1.7, 1.8, 1.9, 2, 2.1, 2.2, 2.3, 2.4, 2.5, 2.6,
2.7, 2.8, 2.9, 3, 3.1, 3.2, 3.3, 3.4, 3.5, 3.6, 3.7, 3.8, 3.9, 4,
4.1, 4.2, 4.3, 4.4, 4.5, 4.6, 4.7, 4.8, 4.9, 5, 5.1, 5.2, 5.3, 5.4,
5.5, 5.6, 5.7, 5.8, 5.9, 6, 6.1, 6.2, 6.3, 6.4, 6.5, 6.6, 6.7, 6.8,
6.9, 7, 7.1, 7.2, 7.3, 7.4, 7.5, 7.6, 7.7, 7.8, 7.9, 8, 8.1, 8.2,
8.3, 8.4, 8.5, 8.6, 8.7, 8.8, 8.9, 9, 9.1, 9.2, 9.3, 9.4, 9.5, 9.6,
9.7, 9.8, 9.9, 10, 10.1, 10.2, 10.3, 10.4, 10.5, 10.6, 10.7, 10.8,
10.9, 11, 11.1, 11.2, 11.3, 11.4, 11.5, 11.6, 11.7, 11.8, 11.9, 12,
12.1, 12.2, 12.3, 12.4, 12.5, 12.6, 12.7, 12.8, 12.9, 13, 13.1,
13.2, 13.3, 13.4, 13.5, 13.6, 13.7, 13.8, 13.9, 14, 14.1, 14.2,
14.3, 14.4, 14.5, 14.6, 14.7, 14.8, 14.9, or 15 points. As a
non-limiting example, a subject's UPDRS-3 score is changed by 0.4
points. As a non-limiting example, a subject's UPDRS-3 score is
changed by 1.6 points. As a non-limiting example, a subject's
UPDRS-3 score is changed by 1.8 points. As a non-limiting example,
a subject's UPDRS-3 score is changed by 8.6 points. As a
non-limiting example, a subject's UPDRS-3 score is changed by 9.6
points.
[0515] A subject's UPDRS-3 (or UPDRS-III) medication score may
increase by 0.1, 0.2, 0.3, 0.4, 0.5, 0.6, 0.7, 0.8, 0.9, 1, 1.1,
1.2, 1.3, 1.4, 1.5, 1.6, 1.7, 1.8, 1.9, 2, 2.1, 2.2, 2.3, 2.4, 2.5,
2.6, 2.7, 2.8, 2.9, 3, 3.1, 3.2, 3.3, 3.4, 3.5, 3.6, 3.7, 3.8, 3.9,
4, 4.1, 4.2, 4.3, 4.4, 4.5, 4.6, 4.7, 4.8, 4.9, 5, 5.1, 5.2, 5.3,
5.4, 5.5, 5.6, 5.7, 5.8, 5.9, 6, 6.1, 6.2, 6.3, 6.4, 6.5, 6.6, 6.7,
6.8, 6.9, 7, 7.1, 7.2, 7.3, 7.4, 7.5, 7.6, 7.7, 7.8, 7.9, 8, 8.1,
8.2, 8.3, 8.4, 8.5, 8.6, 8.7, 8.8, 8.9, 9, 9.1, 9.2, 9.3, 9.4, 9.5,
9.6, 9.7, 9.8, 9.9, 10, 10.1, 10.2, 10.3, 10.4, 10.5, 10.6, 10.7,
10.8, 10.9, 11, 11.1, 11.2, 11.3, 11.4, 11.5, 11.6, 11.7, 11.8,
11.9, 12, 12.1, 12.2, 12.3, 12.4, 12.5, 12.6, 12.7, 12.8, 12.9, 13,
13.1, 13.2, 13.3, 13.4, 13.5, 13.6, 13.7, 13.8, 13.9, 14, 14.1,
14.2, 14.3, 14.4, 14.5, 14.6, 14.7, 14.8, 14.9, or 15 points. As a
non-limiting example, a subject's UPDRS-3 score is increased by 0.4
points. As a non-limiting example, a subject's UPDRS-3 score is
increased by 1.6 points. As a non-limiting example, a subject's
UPDRS-3 score is increased by 1.8 points. As a non-limiting
example, a subject's UPDRS-3 score is increased by 8.6 points. As a
non-limiting example, a subject's UPDRS-3 score is increased by 9.6
points.
[0516] A subject's UPDRS-3 (or UPDRS-III) medication score may
decrease by 0.1, 0.2, 0.3, 0.4, 0.5, 0.6, 0.7, 0.8, 0.9, 1, 1.1,
1.2, 1.3, 1.4, 1.5, 1.6, 1.7, 1.8, 1.9, 2, 2.1, 2.2, 2.3, 2.4, 2.5,
2.6, 2.7, 2.8, 2.9, 3, 3.1, 3.2, 3.3, 3.4, 3.5, 3.6, 3.7, 3.8, 3.9,
4, 4.1, 4.2, 4.3, 4.4, 4.5, 4.6, 4.7, 4.8, 4.9, 5, 5.1, 5.2, 5.3,
5.4, 5.5, 5.6, 5.7, 5.8, 5.9, 6, 6.1, 6.2, 6.3, 6.4, 6.5, 6.6, 6.7,
6.8, 6.9, 7, 7.1, 7.2, 7.3, 7.4, 7.5, 7.6, 7.7, 7.8, 7.9, 8, 8.1,
8.2, 8.3, 8.4, 8.5, 8.6, 8.7, 8.8, 8.9, 9, 9.1, 9.2, 9.3, 9.4, 9.5,
9.6, 9.7, 9.8, 9.9, 10, 10.1, 10.2, 10.3, 10.4, 10.5, 10.6, 10.7,
10.8, 10.9, 11, 11.1, 11.2, 11.3, 11.4, 11.5, 11.6, 11.7, 11.8,
11.9, 12, 12.1, 12.2, 12.3, 12.4, 12.5, 12.6, 12.7, 12.8, 12.9, 13,
13.1, 13.2, 13.3, 13.4, 13.5, 13.6, 13.7, 13.8, 13.9, 14, 14.1,
14.2, 14.3, 14.4, 14.5, 14.6, 14.7, 14.8, 14.9, or 15 points. As a
non-limiting example, a subject's UPDRS-3 score is decreased by 0.4
points. As a non-limiting example, a subject's UPDRS-3 score is
decreased by 1.6 points. As a non-limiting example, a subject's
UPDRS-3 score is decreased by 1.8 points. As a non-limiting
example, a subject's UPDRS-3 score is decrease by 8.6 points. As a
non-limiting example, a subject's UPDRS-3 score is decreased by 9.6
points.
[0517] In one embodiment, a subject's UPDRS-3 (or UPDRS-III)
medication score is reduced by 8.6 at 6 months after administration
of the present disclosure at a dose volume of up to 900 uL per
putamen and a total dose of 4.5.times.10.sup.12 vector genome.
[0518] In one embodiment, a subject's UPDRS-3 (or UPDRS-III)
medication score is reduced by 9.6 at 6 months after administration
of the present disclosure at a dose volume of up to 900 uL per
putamen and a total dose of 1.5.times.10.sup.12 vector genome.
[0519] In one embodiment, a subject's UPDRS-3 (or UPDRS-III)
medication score is reduced by 9.6 at 12 months after
administration of the present disclosure at a dose volume of up to
900 uL per putamen and a total dose of 1.5.times.10.sup.12 vector
genome.
[0520] In one embodiment, a subject's average amount of Parkinson's
disease medication was about 1500 mg per day prior to
administration of the present disclosure. As a non-limiting
example, the Parkinson's disease medication is levodopa.
[0521] In one embodiment, a subject's UPDRS-II score is evaluated
prior to administration of the present disclosure. The UPDRS-II
score of a subject prior to administration of the present
disclosure is between 20 and 50, such as, but not limited to, 20,
21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37,
38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, and 50.
[0522] A subject's UPDRS-2 (or UPDRS-II) score may change by 0.1,
0.2, 0.3, 0.4, 0.5, 0.6, 0.7, 0.8, 0.9, 1, 1.1, 1.2, 1.3, 1.4, 1.5,
1.6, 1.7, 1.8, 1.9, 2, 2.1, 2.2, 2.3, 2.4, 2.5, 2.6, 2.7, 2.8, 2.9,
3, 3.1, 3.2, 3.3, 3.4, 3.5, 3.6, 3.7, 3.8, 3.9, 4, 4.1, 4.2, 4.3,
4.4, 4.5, 4.6, 4.7, 4.8, 4.9, 5, 5.1, 5.2, 5.3, 5.4, 5.5, 5.6, 5.7,
5.8, 5.9, 6, 6.1, 6.2, 6.3, 6.4, 6.5, 6.6, 6.7, 6.8, 6.9, 7, 7.1,
7.2, 7.3, 7.4, 7.5, 7.6, 7.7, 7.8, 7.9, 8, 8.1, 8.2, 8.3, 8.4, 8.5,
8.6, 8.7, 8.8, 8.9, 9, 9.1, 9.2, 9.3, 9.4, 9.5, 9.6, 9.7, 9.8, 9.9,
10, 10.1, 10.2, 10.3, 10.4, 10.5, 10.6, 10.7, 10.8, 10.9, 11, 11.1,
11.2, 11.3, 11.4, 11.5, 11.6, 11.7, 11.8, 11.9, 12, 12.1, 12.2,
12.3, 12.4, 12.5, 12.6, 12.7, 12.8, 12.9, 13, 13.1, 13.2, 13.3,
13.4, 13.5, 13.6, 13.7, 13.8, 13.9, 14, 14.1, 14.2, 14.3, 14.4,
14.5, 14.6, 14.7, 14.8, 14.9, or 15 points.
[0523] A subject's UPDRS-2 (or UPDRS-II) score may increase by 0.1,
0.2, 0.3, 0.4, 0.5, 0.6, 0.7, 0.8, 0.9, 1, 1.1, 1.2, 1.3, 1.4, 1.5,
1.6, 1.7, 1.8, 1.9, 2, 2.1, 2.2, 2.3, 2.4, 2.5, 2.6, 2.7, 2.8, 2.9,
3, 3.1, 3.2, 3.3, 3.4, 3.5, 3.6, 3.7, 3.8, 3.9, 4, 4.1, 4.2, 4.3,
4.4, 4.5, 4.6, 4.7, 4.8, 4.9, 5, 5.1, 5.2, 5.3, 5.4, 5.5, 5.6, 5.7,
5.8, 5.9, 6, 6.1, 6.2, 6.3, 6.4, 6.5, 6.6, 6.7, 6.8, 6.9, 7, 7.1,
7.2, 7.3, 7.4, 7.5, 7.6, 7.7, 7.8, 7.9, 8, 8.1, 8.2, 8.3, 8.4, 8.5,
8.6, 8.7, 8.8, 8.9, 9, 9.1, 9.2, 9.3, 9.4, 9.5, 9.6, 9.7, 9.8, 9.9,
10, 10.1, 10.2, 10.3, 10.4, 10.5, 10.6, 10.7, 10.8, 10.9, 11, 11.1,
11.2, 11.3, 11.4, 11.5, 11.6, 11.7, 11.8, 11.9, 12, 12.1, 12.2,
12.3, 12.4, 12.5, 12.6, 12.7, 12.8, 12.9, 13, 13.1, 13.2, 13.3,
13.4, 13.5, 13.6, 13.7, 13.8, 13.9, 14, 14.1, 14.2, 14.3, 14.4,
14.5, 14.6, 14.7, 14.8, 14.9, or 15 points.
[0524] A subject's UPDRS-2 (or UPDRS-II) score may decrease by 0.1,
0.2, 0.3, 0.4, 0.5, 0.6, 0.7, 0.8, 0.9, 1, 1.1, 1.2, 1.3, 1.4, 1.5,
1.6, 1.7, 1.8, 1.9, 2, 2.1, 2.2, 2.3, 2.4, 2.5, 2.6, 2.7, 2.8, 2.9,
3, 3.1, 3.2, 3.3, 3.4, 3.5, 3.6, 3.7, 3.8, 3.9, 4, 4.1, 4.2, 4.3,
4.4, 4.5, 4.6, 4.7, 4.8, 4.9, 5, 5.1, 5.2, 5.3, 5.4, 5.5, 5.6, 5.7,
5.8, 5.9, 6, 6.1, 6.2, 6.3, 6.4, 6.5, 6.6, 6.7, 6.8, 6.9, 7, 7.1,
7.2, 7.3, 7.4, 7.5, 7.6, 7.7, 7.8, 7.9, 8, 8.1, 8.2, 8.3, 8.4, 8.5,
8.6, 8.7, 8.8, 8.9, 9, 9.1, 9.2, 9.3, 9.4, 9.5, 9.6, 9.7, 9.8, 9.9,
10, 10.1, 10.2, 10.3, 10.4, 10.5, 10.6, 10.7, 10.8, 10.9, 11, 11.1,
11.2, 11.3, 11.4, 11.5, 11.6, 11.7, 11.8, 11.9, 12, 12.1, 12.2,
12.3, 12.4, 12.5, 12.6, 12.7, 12.8, 12.9, 13, 13.1, 13.2, 13.3,
13.4, 13.5, 13.6, 13.7, 13.8, 13.9, 14, 14.1, 14.2, 14.3, 14.4,
14.5, 14.6, 14.7, 14.8, 14.9, or 15 points. As a non-limiting
example, a 3.6 point reduction is seen 12 months after
administration with the present disclosure. As a non-limiting
example, a 3.6 point reduction is seen 6 months after
administration with the present disclosure.
[0525] In one embodiment, a subject's UPDRS-II score is decreased
by 2 to 4 points after administration of the present disclosure as
compared to the UPDRS-II score prior to administration.
[0526] In one embodiment, a subject's UPDRS-II score is decreased
by 2 to 3 points 6 months after administration of the present
disclosure at a dose volume of up to 900 uL per putamen and a total
dose of 1.5.times.10.sup.12 vector genome, as compared to the
UPDRS-II medication score prior to administration.
[0527] In one embodiment, a subject's UPDRS-II score is decreased
by 2 to 3 points 12 months after administration of the present
disclosure at a dose volume of up to 900 uL per putamen and a total
dose of 1.5.times.10.sup.12 vector genome, as compared to the
UPDRS-II medication score prior to administration.
[0528] In one embodiment, a subject's UPDRS-II score is decreased
by 3 to 4 points 6 months after administration of the present
disclosure at a dose volume of up to 900 uL per putamen and a total
dose of 4.5.times.10.sup.12 vector genome, as compared to the
UPDRS-II medication score prior to administration.
[0529] In one embodiment, the present disclosure is used to improve
a subject's motor function.
[0530] In one embodiment, the present disclosure is used to control
a subject's motor function and improve their quality of life.
[0531] In one embodiment, the present disclosure is used to reduce
the dosage of Parkinson's medication a subject needs to take to
improve a subject's motor function.
[0532] In one embodiment, a single administration of the present
disclosure into the putamen of a subject provides improved motor
function as compared to motor function prior to treatment.
[0533] In one embodiment, a single administration of the present
disclosure in to the putamen of a subject provides improved motor
function and a reduction in the amount of levodopa the subject
requires to manage symptoms.
[0534] In one embodiment, a single administration of the present
disclosure in to the putamen of a subject provides improved motor
function and a reduction in the amount of dopaminergic medication
the subject requires to manage symptoms.
[0535] In one embodiment, the amount the daily dose of Parkinson's
medication (e.g., Levodopa) is reduced by 10-50% after
administration of the present disclosure. As a non-limiting
example, the reduction is seen 6 months after administration of the
present disclosure. As a non-limiting example, the reduction is
seen 12 months after administration of the present disclosure.
[0536] In one embodiment, the amount the daily dose of Parkinson's
medication (e.g., Levodopa) is reduced by 10-20% after
administration of the present disclosure. As a non-limiting
example, the reduction of Parkinson's medication (e.g., Levodopa)
is 10%. As a non-limiting example, the reduction of Parkinson's
medication (e.g., Levodopa) is 11%. As a non-limiting example, the
reduction of Parkinson's medication (e.g., Levodopa) is 12%. As a
non-limiting example, the reduction of Parkinson's medication
(e.g., Levodopa) is 13%. As a non-limiting example, the reduction
of Parkinson's medication (e.g., Levodopa) is 14%. As a
non-limiting example, the reduction of Parkinson's medication
(e.g., Levodopa) is 15%. As a non-limiting example, the reduction
of Parkinson's medication (e.g., Levodopa) is 16%. As a
non-limiting example, the reduction of Parkinson's medication
(e.g., Levodopa) is 17%. As a non-limiting example, the reduction
of Parkinson's medication (e.g., Levodopa) is 18%. As a
non-limiting example, the reduction of Parkinson's medication
(e.g., Levodopa) is 19%. As a non-limiting example, the reduction
of Parkinson's medication (e.g., Levodopa) is 20%.
[0537] In one embodiment, a 14% reduction in the amount of the
daily dose of Parkinson's medication (e.g., Levodopa) required by a
subject to manage symptoms occurs 6 months after administration of
the present disclosure at a dose volume of up to 450 uL per putamen
and a total dose of 7.5.times.10.sup.11 vector genome.
[0538] In one embodiment, a 10% reduction in the amount of the
daily dose of Parkinson's medication (e.g., Levodopa) required by a
subject to manage symptoms occurs 12 months after administration of
the present disclosure at a dose volume of up to 450 uL per putamen
and a total dose of 7.5.times.10.sup.11 vector genome.
[0539] In one embodiment, the amount the daily dose of Parkinson's
medication (e.g., Levodopa) is reduced by 20-30% after
administration of the present disclosure. As a non-limiting
example, the reduction of Parkinson's medication (e.g., Levodopa)
is 20%. As a non-limiting example, the reduction of Parkinson's
medication (e.g., Levodopa) is 21%. As a non-limiting example, the
reduction of Parkinson's medication (e.g., Levodopa) is 22%. As a
non-limiting example, the reduction of Parkinson's medication
(e.g., Levodopa) is 23%. As a non-limiting example, the reduction
of Parkinson's medication (e.g., Levodopa) is 24%. As a
non-limiting example, the reduction of Parkinson's medication
(e.g., Levodopa) is 25%. As a non-limiting example, the reduction
of Parkinson's medication (e.g., Levodopa) is 26%. As a
non-limiting example, the reduction of Parkinson's medication
(e.g., Levodopa) is 27%. As a non-limiting example, the reduction
of Parkinson's medication (e.g., Levodopa) is 28%. As a
non-limiting example, the reduction of Parkinson's medication
(e.g., Levodopa) is 29%. As a non-limiting example, the reduction
of Parkinson's medication (e.g., Levodopa) is 30%.
[0540] In one embodiment, a 27% reduction in the amount of the
daily dose of Parkinson's medication (e.g., Levodopa) required by a
subject to manage symptoms occurs 12 months after administration of
the present disclosure at a dose volume of up to 900 uL per putamen
and a total dose of 1.5.times.10.sup.12 vector genome.
[0541] In one embodiment, a 28% reduction in the amount of the
daily dose of Parkinson's medication (e.g., Levodopa) required by a
subject to manage symptoms occurs 12 months after administration of
the present disclosure at a dose volume of up to 900 uL per putamen
and a total dose of 1.5.times.10.sup.12 vector genome.
[0542] In one embodiment, a 29% reduction in the amount of the
daily dose of Parkinson's medication (e.g., Levodopa) required by a
subject to manage symptoms occurs 12 months after administration of
the present disclosure at a dose volume of up to 900 uL per putamen
and a total dose of 1.5.times.10.sup.12 vector genome.
[0543] In one embodiment, the amount the daily dose of Parkinson's
medication (e.g., Levodopa) is reduced by 30-40% after
administration of the present disclosure. As a non-limiting
example, the reduction of Parkinson's medication (e.g., Levodopa)
is 30%. As a non-limiting example, the reduction of Parkinson's
medication (e.g., Levodopa) is 31%. As a non-limiting example, the
reduction of Parkinson's medication (e.g., Levodopa) is 32%. As a
non-limiting example, the reduction of Parkinson's medication
(e.g., Levodopa) is 33%. As a non-limiting example, the reduction
of Parkinson's medication (e.g., Levodopa) is 34%. As a
non-limiting example, the reduction of Parkinson's medication
(e.g., Levodopa) is 35%. As a non-limiting example, the reduction
of Parkinson's medication (e.g., Levodopa) is 36%. As a
non-limiting example, the reduction of Parkinson's medication
(e.g., Levodopa) is 37%. As a non-limiting example, the reduction
of Parkinson's medication (e.g., Levodopa) is 38%. As a
non-limiting example, the reduction of Parkinson's medication
(e.g., Levodopa) is 39%. As a non-limiting example, the reduction
of Parkinson's medication (e.g., Levodopa) is 40%.
[0544] In one embodiment, a 34% reduction in the amount of the
daily dose of Parkinson's medication (e.g., Levodopa) required by a
subject to manage symptoms occurs 6 months after administration of
the present disclosure at a dose volume of up to 900 uL per putamen
and a total dose of 1.5.times.10.sup.12 vector genome.
[0545] In one embodiment, the amount the daily dose of Parkinson's
medication (e.g., Levodopa) is reduced by 40-50% after
administration of the present disclosure. As a non-limiting
example, the reduction of Parkinson's medication (e.g., Levodopa)
is 40%. As a non-limiting example, the reduction of Parkinson's
medication (e.g., Levodopa) is 41%. As a non-limiting example, the
reduction of Parkinson's medication (e.g., Levodopa) is 42%. As a
non-limiting example, the reduction of Parkinson's medication
(e.g., Levodopa) is 43%. As a non-limiting example, the reduction
of Parkinson's medication (e.g., Levodopa) is 44%. As a
non-limiting example, the reduction of Parkinson's medication
(e.g., Levodopa) is 45%. As a non-limiting example, the reduction
of Parkinson's medication (e.g., Levodopa) is 46%. As a
non-limiting example, the reduction of Parkinson's medication
(e.g., Levodopa) is 47%. As a non-limiting example, the reduction
of Parkinson's medication (e.g., Levodopa) is 48%. As a
non-limiting example, the reduction of Parkinson's medication
(e.g., Levodopa) is 49%. As a non-limiting example, the reduction
of Parkinson's medication (e.g., Levodopa) is 50%.
[0546] In one embodiment, a 34% reduction in the amount of the
daily dose of Parkinson's medication (e.g., Levodopa) required by a
subject to manage symptoms occurs 6 months after administration of
the present disclosure at a dose volume of up to 900 uL per putamen
and a total dose of 4.5.times.10.sup.12 vector genome.
[0547] In one embodiment, the amount the daily dose of Parkinson's
medication (e.g., Levodopa) is reduced by 108-641 mg after
administration of the present disclosure. As a non-limiting
example, the reduction is seen 6 months after administration of the
present disclosure. As a non-limiting example, the reduction is
seen 12 months after administration of the present disclosure.
[0548] In one embodiment, the amount the daily dose of Parkinson's
medication (e.g., Levodopa) is reduced by 108-339 mg after
administration of the present disclosure. As a non-limiting
example, the reduction of Parkinson's medication (e.g., Levodopa)
is 108 mg. As a non-limiting example, the reduction of Parkinson's
medication (e.g., Levodopa) is 134 mg. As a non-limiting example,
the reduction of Parkinson's medication (e.g., Levodopa) is 159 mg.
As a non-limiting example, the reduction of Parkinson's medication
(e.g., Levodopa) is 154 mg. As a non-limiting example, the
reduction of Parkinson's medication (e.g., Levodopa) is 208 mg. As
a non-limiting example, the reduction of Parkinson's medication
(e.g., Levodopa) is 231 mg. As a non-limiting example, the
reduction of Parkinson's medication (e.g., Levodopa) is 254 mg. As
a non-limiting example, the reduction of Parkinson's medication
(e.g., Levodopa) is 276 mg. As a non-limiting example, the
reduction of Parkinson's medication (e.g., Levodopa) is 298 mg. As
a non-limiting example, the reduction of Parkinson's medication
(e.g., Levodopa) is 319 mg. As a non-limiting example, the
reduction of Parkinson's medication (e.g., Levodopa) is 339 mg.
[0549] In one embodiment, a 208 mg reduction in the amount of the
daily dose of Parkinson's medication (e.g., Levodopa) required by a
subject to manage symptoms occurs 6 months after administration of
the present disclosure at a dose volume of up to 450 uL per putamen
and a total dose of 7.5.times.10.sup.11 vector genome.
[0550] In one embodiment, a 108 mg reduction in the amount of the
daily dose of Parkinson's medication (e.g., Levodopa) required by a
subject to manage symptoms occurs 12 months after administration of
the present disclosure at a dose volume of up to 450 uL per putamen
and a total dose of 7.5.times.10.sup.11 vector genome.
[0551] In one embodiment, the amount the daily dose of Parkinson's
medication (e.g., Levodopa) is reduced by 339-505 mg after
administration of the present disclosure. As a non-limiting
example, the reduction of Parkinson's medication (e.g., Levodopa)
is 339 mg. As a non-limiting example, the reduction of Parkinson's
medication (e.g., Levodopa) is 358 mg. As a non-limiting example,
the reduction of Parkinson's medication (e.g., Levodopa) is 377 mg.
As a non-limiting example, the reduction of Parkinson's medication
(e.g., Levodopa) is 396 mg. As a non-limiting example, the
reduction of Parkinson's medication (e.g., Levodopa) is 413 mg. As
a non-limiting example, the reduction of Parkinson's medication
(e.g., Levodopa) is 430 mg. As a non-limiting example, the
reduction of Parkinson's medication (e.g., Levodopa) is 446 mg. As
a non-limiting example, the reduction of Parkinson's medication
(e.g., Levodopa) is 462 mg. As a non-limiting example, the
reduction of Parkinson's medication (e.g., Levodopa) is 477 mg. As
a non-limiting example, the reduction of Parkinson's medication
(e.g., Levodopa) is 491 mg. As a non-limiting example, the
reduction of Parkinson's medication (e.g., Levodopa) is 505 mg.
[0552] In one embodiment, a 462 mg reduction in the amount of the
daily dose of Parkinson's medication (e.g., Levodopa) required by a
subject to manage symptoms occurs 12 months after administration of
the present disclosure at a dose volume of up to 900 uL per putamen
and a total dose of 1.5.times.10.sup.12 vector genome.
[0553] In one embodiment, a 477 mg reduction in the amount of the
daily dose of Parkinson's medication (e.g., Levodopa) required by a
subject to manage symptoms occurs 12 months after administration of
the present disclosure at a dose volume of up to 900 uL per putamen
and a total dose of 1.5.times.10.sup.12 vector genome.
[0554] In one embodiment, a 491 mg reduction in the amount of the
daily dose of Parkinson's medication (e.g., Levodopa) required by a
subject to manage symptoms occurs 12 months after administration of
the present disclosure at a dose volume of up to 900 uL per putamen
and a total dose of 1.5.times.10.sup.12 vector genome.
[0555] In one embodiment, the amount the daily dose of Parkinson's
medication (e.g., Levodopa) is reduced by 505-606 mg after
administration of the present disclosure. As a non-limiting
example, the reduction of Parkinson's medication (e.g., Levodopa)
is 505 mg. As a non-limiting example, the reduction of Parkinson's
medication (e.g., Levodopa) is 518 mg. As a non-limiting example,
the reduction of Parkinson's medication (e.g., Levodopa) is 530 mg.
As a non-limiting example, the reduction of Parkinson's medication
(e.g., Levodopa) is 542 mg. As a non-limiting example, the
reduction of Parkinson's medication (e.g., Levodopa) is 553 mg. As
a non-limiting example, the reduction of Parkinson's medication
(e.g., Levodopa) is 563 mg. As a non-limiting example, the
reduction of Parkinson's medication (e.g., Levodopa) is 573 mg. As
a non-limiting example, the reduction of Parkinson's medication
(e.g., Levodopa) is 582 mg. As a non-limiting example, the
reduction of Parkinson's medication (e.g., Levodopa) is 591 mg. As
a non-limiting example, the reduction of Parkinson's medication
(e.g., Levodopa) is 599 mg. As a non-limiting example, the
reduction of Parkinson's medication (e.g., Levodopa) is 606 mg.
[0556] In one embodiment, a 553 mg reduction in the amount of the
daily dose of Parkinson's medication (e.g., Levodopa) required by a
subject to manage symptoms occurs 6 months after administration of
the present disclosure at a dose volume of up to 900 uL per putamen
and a total dose of 1.5.times.10.sup.12 vector genome.
[0557] In one embodiment, the amount the daily dose of Parkinson's
medication (e.g., Levodopa) is reduced by 606-641 mg after
administration of the present disclosure. As a non-limiting
example, the reduction of Parkinson's medication (e.g., Levodopa)
is 606 mg. As a non-limiting example, the reduction of Parkinson's
medication (e.g., Levodopa) is 612 mg. As a non-limiting example,
the reduction of Parkinson's medication (e.g., Levodopa) is 618 mg.
As a non-limiting example, the reduction of Parkinson's medication
(e.g., Levodopa) is 623 mg. As a non-limiting example, the
reduction of Parkinson's medication (e.g., Levodopa) is 628 mg. As
a non-limiting example, the reduction of Parkinson's medication
(e.g., Levodopa) is 632 mg. As a non-limiting example, the
reduction of Parkinson's medication (e.g., Levodopa) is 635 mg. As
a non-limiting example, the reduction of Parkinson's medication
(e.g., Levodopa) is 637 mg. As a non-limiting example, the
reduction of Parkinson's medication (e.g., Levodopa) is 639 mg. As
a non-limiting example, the reduction of Parkinson's medication
(e.g., Levodopa) is 641 mg.
[0558] In one embodiment, a 553 mg reduction in the amount of the
daily dose of Parkinson's medication (e.g., Levodopa) required by a
subject to manage symptoms occurs 6 months after administration of
the present disclosure at a dose volume of up to 900 uL per putamen
and a total dose of 4.5.times.10.sup.12 vector genome.
[0559] In one embodiment, the putaminal AADC enzyme activity is
increased in a subject after administration with the present
disclosure. As a non-limiting example, the increase is seen for at
least 6 months relative to the baseline.
[0560] In one embodiment, the putaminal AADC enzyme activity is
increased by 10-20% in a subject after administration of the
present disclosure. As a non-limiting example, the increase in
putaminal AADC enzyme activity is about 10%. As a non-limiting
example, the increase in putaminal AADC enzyme activity is about
11%. As a non-limiting example, the increase in putaminal AADC
enzyme activity is about 12%. As a non-limiting example, the
increase in putaminal AADC enzyme activity is about 13%. As a
non-limiting example, the increase in putaminal AADC enzyme
activity is about 14%. As a non-limiting example, the increase in
putaminal AADC enzyme activity is about 15%. As a non-limiting
example, the increase in putaminal AADC enzyme activity is about
16%. As a non-limiting example, the increase in putaminal AADC
enzyme activity is about 17%. As a non-limiting example, the
increase in putaminal AADC enzyme activity is about 18%. As a
non-limiting example, the increase in putaminal AADC enzyme
activity is about 19%. As a non-limiting example, the increase in
putaminal AADC enzyme activity is about 20%.
[0561] In one embodiment, the putaminal AADC enzyme activity is
increased by about 13% in a subject after administration of the
present disclosure at a dose volume of up to 450 uL per putamen and
a total dose of 7.5.times.10.sup.11 vector genome.
[0562] In one embodiment, the putaminal AADC enzyme activity is
increased by 50-60% in a subject after administration of the
present disclosure. As a non-limiting example, the increase in
putaminal AADC enzyme activity is about 50%. As a non-limiting
example, the increase in putaminal AADC enzyme activity is about
51%. As a non-limiting example, the increase in putaminal AADC
enzyme activity is about 52%. As a non-limiting example, the
increase in putaminal AADC enzyme activity is about 53%. As a
non-limiting example, the increase in putaminal AADC enzyme
activity is about 54%. As a non-limiting example, the increase in
putaminal AADC enzyme activity is about 55%. As a non-limiting
example, the increase in putaminal AADC enzyme activity is about
56%. As a non-limiting example, the increase in putaminal AADC
enzyme activity is about 57%. As a non-limiting example, the
increase in putaminal AADC enzyme activity is about 58%. As a
non-limiting example, the increase in putaminal AADC enzyme
activity is about 59%. As a non-limiting example, the increase in
putaminal AADC enzyme activity is about 60%.
[0563] In one embodiment, the putaminal AADC enzyme activity is
increased by about 56% in a subject after administration of the
present disclosure at a dose volume of up to 900 uL per putamen and
a total dose of 1.5.times.10.sup.12 vector genome.
[0564] In one embodiment, the putaminal AADC enzyme activity is
increased by 70-85% in a subject after administration of the
present disclosure. As a non-limiting example, the increase in
putaminal AADC enzyme activity is about 70%. As a non-limiting
example, the increase in putaminal AADC enzyme activity is about
71%. As a non-limiting example, the increase in putaminal AADC
enzyme activity is about 72%. As a non-limiting example, the
increase in putaminal AADC enzyme activity is about 73%. As a
non-limiting example, the increase in putaminal AADC enzyme
activity is about 74%. As a non-limiting example, the increase in
putaminal AADC enzyme activity is about 75%. As a non-limiting
example, the increase in putaminal AADC enzyme activity is about
76%. As a non-limiting example, the increase in putaminal AADC
enzyme activity is about 77%. As a non-limiting example, the
increase in putaminal AADC enzyme activity is about 78%. As a
non-limiting example, the increase in putaminal AADC enzyme
activity is about 79%. As a non-limiting example, the increase in
putaminal AADC enzyme activity is about 80%. As a non-limiting
example, the increase in putaminal AADC enzyme activity is about
81%. As a non-limiting example, the increase in putaminal AADC
enzyme activity is about 82%. As a non-limiting example, the
increase in putaminal AADC enzyme activity is about 83%. As a
non-limiting example, the increase in putaminal AADC enzyme
activity is about 84%. As a non-limiting example, the increase in
putaminal AADC enzyme activity is about 85%.
[0565] In one embodiment, the putaminal AADC enzyme activity is
increased by about 79% in a subject after administration of the
present disclosure at a dose volume of up to 900 uL per putamen and
a total dose of 4.5.times.10.sup.12 vector genome.
[0566] In one embodiment, the dopamine level of a subject increased
after administration of the present disclosure. As a non-limiting
example, the amount of dopamine increased by 10-20%, 15-25%,
20-30%, 25-35%, 30-40%, 35-45%, 40-50%, 45-55%, 50-60%, 55-65%,
60-70%, 65-75%, 70-80%, 75-85%, 80-90%, 85-95%, 90-100%, or
95-100%.
Circadian Rhythm and Sleep-Wake Cycles
[0567] Circadian rhythms are physical, mental and behavioral
changes that tend to follow a 24 hour cycle. Circadian rhythms can
influence sleep-wake cycles, hormone release, body temperature and
other bodily functions. Changes in the circadian rhythm can cause
conditions and/or disorder such as, but not limited to sleep
disorders (e.g., insomnia), depression, bipolar disorder, seasonal
affective disorder, obesity and diabetes.
[0568] In one embodiment, the AAV particles described herein may be
used to treat insomnia.
[0569] The sleep-wake cycle comprises periods of sleep and periods
of wake. Generally, in a 24 hour period the total hours of sleep
are less than the total hours of wakefulness. As a non-limiting
example, the sleep-wake cycle comprises 7-9 hours of sleep and
15-17 hours of wakefulness. As a non-limiting example, the
sleep-wake cycle comprises 8 hours of sleep and 16 hours of
wakefulness. As a non-limiting example, the sleep-wake cycle
comprises 8-10 hours of sleep and 14-16 hours of wakefulness.
[0570] In one embodiment, the sleep-wake cycle of a subject is
improved by administering to the subject the AAV particles
described herein.
[0571] In one embodiment, the sleep-wake cycle of a subject is
regulated by administering to the subject the AAV particles
described herein. As a non-limiting example, the regulation may be
the correction of more periods of sleep occurring at night and less
periods of sleep occurring
[0572] In one embodiment, the sleep-wake cycle of a subject
administered the AAV particles described herein improves as
compared to the sleep-wake cycle of the subject prior to
administration of the AAV particles. As a non-limiting example, the
subject has an increased period of sleep and a decreased period of
wakefulness. As another non-limiting example, the subject has a
decreased period of sleep and an increased period of
wakefulness.
[0573] In one embodiment, the sleep-wake cycle of a subject
administered the AAV particles described herein is regulated as
compared to the sleep-wake cycle of the subject prior to
administration of the AAV particles. As a non-limiting example, the
length of the periods of sleep and the periods of wakefulness may
be about the same (e.g., +/-1 hour) for at least 2 days. As another
non-limiting example, the length of the periods of sleep and the
periods of wakefulness if a 24 hours period may be within 10
minutes, 15 minutes, 20 minutes, 25 minutes, 30 minutes, 35
minutes, 40 minutes, 45 minutes, 50 minutes, 55 minutes, 1 hour,
1.5 hours, or 2 hours of the previous 24 hour period.
[0574] In one embodiment, the amount of rapid eye movement (REM)
sleep a subject experiences in a 24 hour period is altered after
the subject is administered the AAV particles described herein. REM
sleep is generally considered an active period of sleep marked by
intense brain activity where brain waves are fast and
desynchronized. An adult, on average, spends about 20-25% of their
total daily sleep period in REM sleep. As a non-limiting example,
the amount of REM sleep is decreased by 1%, 2%, 3%, 4%, 5%, 6%, 7%,
8%, 9%, 10%, 11%, 12%, 13%, 14%, 15%, 20%, 25%, 30%, 35%, 40%, 45%,
50%, 55%, 60%, 65% or more than 65%. As a non-limiting example, the
amount of REM sleep is decreased by 1-10%, 5-10%, 5-15%, 10-15%,
15-20%, 15-25%, 20-25%, 20-30%, 25-30%, 25-35%, 30-35%, 30-40%,
35-40%, 40-50% or 40-60%. As a non-limiting example, the amount of
REM sleep is increased by 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%,
11%, 12%, 13%, 14%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%,
60%, 65% or more than 65%. As a non-limiting example, the amount of
REM sleep is increased by 1-5%, 1-10%, 5-10%, 5-15%, 10-15%,
15-20%, 15-25%, 20-25%, 20-30%, 25-30%, 25-35%, 30-35%, 30-40%,
35-40%, 40-50% or 40-60%.
[0575] In one embodiment, the amount of non-REM (NREM) sleep a
subject experiences in a 24 hour period is altered after the
subject is administered the AAV particles described herein. NREM
sleep is generally characterized by a reduction in physiological
activity since as the brain waves, measured by EEG, get slower and
have greater amplitude. NREM has four stages: Stage 1 is the time
of drowsiness or transition from being awake to falling asleep
where the brain waves and muscle activity begin to slow; Stage 2 is
a period of light sleep during which eye movements stop and brain
waves become slower with occasional bursts of rapid waves
(sometimes called sleep spindles); Stage 3 and Stage 4
(collectively referred to as slow wave sleep) are characterized by
the presence of slow brain waves (delta waves) interspersed with
smaller faster waves where there are no eye movements. An adult, on
average, spends about 75-80% of their total daily sleep period in
NREM sleep with about half of their total daily sleep time in NREM
stage 2 sleep.
[0576] In one embodiment, the amount of NREM sleep a subject
experiences is altered after the subject is administered the AAV
particles described herein. As a non-limiting example, the amount
of NREM sleep is decreased by 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%,
10%, 11%, 12%, 13%, 14%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%,
55%, 60%, 65% or more than 65%. As a non-limiting example, the
amount of NREM sleep is decreased by 1-10%, 5-10%, 5-15%, 10-15%,
15-20%, 15-25%, 20-25%, 20-30%, 25-30%, 25-35%, 30-35%, 30-40%,
35-40%, 40-50% or 40-60%. As a non-limiting example, the amount of
NREM sleep is increased by 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%,
11%, 12%, 13%, 14%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%,
60%, 65% or more than 65%. As a non-limiting example, the amount of
NREM sleep is increased by 1-5%, 1-10%, 5-10%, 5-15%, 10-15%,
15-20%, 15-25%, 20-25%, 20-30%, 25-30%, 25-35%, 30-35%, 30-40%,
35-40%, 40-50% or 40-60%.
[0577] In one embodiment, the amount of NREM Stage 1 sleep a
subject experiences is altered after the subject is administered
the AAV particles described herein. As a non-limiting example, the
amount of NREM Stage 1 sleep is decreased by 1%, 2%, 3%, 4%, 5%,
6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, 15%, 20%, 25%, 30%, 35%,
40%, 45%, 50%, 55%, 60%, 65% or more than 65%. As a non-limiting
example, the amount of NREM Stage 1 sleep is decreased by 1-10%,
5-10%, 5-15%, 10-15%, 15-20%, 15-25%, 20-25%, 20-30%, 25-30%,
25-35%, 30-35%, 30-40%, 35-40%, 40-50% or 40-60%. As a non-limiting
example, the amount of NREM Stage 1 sleep is increased by 1%, 2%,
3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, 15%, 20%, 25%,
30%, 35%, 40%, 45%, 50%, 55%, 60%, 65% or more than 65%. As a
non-limiting example, the amount of NREM Stage 1 sleep is increased
by 1-5%, 1-10%, 5-10%, 5-15%, 10-15%, 15-20%, 15-25%, 20-25%,
20-30%, 25-30%, 25-35%, 30-35%, 30-40%, 35-40%, 40-50% or
40-60%.
[0578] In one embodiment, the amount of NREM Stage 2 sleep a
subject experiences is altered after the subject is administered
the AAV particles described herein. As a non-limiting example, the
amount of NREM Stage 2 sleep is decreased by 1%, 2%, 3%, 4%, 5%,
6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, 15%, 20%, 25%, 30%, 35%,
40%, 45%, 50%, 55%, 60%, 65% or more than 65%. As a non-limiting
example, the amount of NREM Stage 2 sleep is decreased by 1-10%,
5-10%, 5-15%, 10-15%, 15-20%, 15-25%, 20-25%, 20-30%, 25-30%,
25-35%, 30-35%, 30-40%, 35-40%, 40-50% or 40-60%. As a non-limiting
example, the amount of NREM Stage 2 sleep is increased by 1%, 2%,
3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, 15%, 20%, 25%,
30%, 35%, 40%, 45%, 50%, 55%, 60%, 65% or more than 65%. As a
non-limiting example, the amount of NREM Stage 2 sleep is increased
by 1-5%, 1-10%, 5-10%, 5-15%, 10-15%, 15-20%, 15-25%, 20-25%,
20-30%, 25-30%, 25-35%, 30-35%, 30-40%, 35-40%, 40-50% or
40-60%.
[0579] In one embodiment, the amount of NREM Stage 3 and 4 sleep a
subject experiences is altered after the subject is administered
the AAV particles described herein. As a non-limiting example, the
amount of NREM Stage 3 and 4 sleep is decreased by 1%, 2%, 3%, 4%,
5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, 15%, 20%, 25%, 30%,
35%, 40%, 45%, 50%, 55%, 60%, 65% or more than 65%. As a
non-limiting example, the amount of NREM Stage 3 and 4 sleep is
decreased by 1-10%, 5-10%, 5-15%, 10-15%, 15-20%, 15-25%, 20-25%,
20-30%, 25-30%, 25-35%, 30-35%, 30-40%, 35-40%, 40-50% or 40-60%.
As a non-limiting example, the amount of NREM Stage 3 and 4 sleep
is increased by 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%,
13%, 14%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65% or
more than 65%. As a non-limiting example, the amount of NREM Stage
3 and 4 sleep is increased by 1-5%, 1-10%, 5-10%, 5-15%, 10-15%,
15-20%, 15-25%, 20-25%, 20-30%, 25-30%, 25-35%, 30-35%, 30-40%,
35-40%, 40-50% or 40-60%.
[0580] In one embodiment, periods of NREM and REM cycles are more
consistent in a subject after the subject is administered the AAV
particles described herein. Generally NREM and REM cycles alternate
every 90 to 110 minutes four to six times per night.
V. Kits and Devices
Kits
[0581] In one embodiment, the disclosure provides a variety of kits
for conveniently and/or effectively carrying out methods of the
present disclosure. Typically kits will comprise sufficient amounts
and/or numbers of components to allow a user to perform multiple
treatments of a subject(s) and/or to perform multiple
experiments.
[0582] Any of the AAV particles of the present disclosure may be
comprised in a kit. In some embodiments, kits may further include
reagents and/or instructions for creating and/or synthesizing
compounds and/or compositions of the present disclosure. In some
embodiments, kits may also include one or more buffers. In some
embodiments, kits of the disclosure may include components for
making protein or nucleic acid arrays or libraries and thus, may
include, for example, solid supports.
[0583] In some embodiments, kit components may be packaged either
in aqueous media or in lyophilized form. The container means of the
kits will generally include at least one vial, test tube, flask,
bottle, syringe or other container means, into which a component
may be placed, and preferably, suitably aliquoted. Where there is
more than one kit component, (labeling reagent and label may be
packaged together), kits may also generally contain second, third
or other additional containers into which additional components may
be separately placed. In some embodiments, kits may also comprise
second container means for containing sterile, pharmaceutically
acceptable buffers and/or other diluents. In some embodiments,
various combinations of components may be comprised in one or more
vial. Kits of the present disclosure may also typically include
means for containing compounds and/or compositions of the present
disclosure, e.g., proteins, nucleic acids, and any other reagent
containers in close confinement for commercial sale. Such
containers may include injection or blow-molded plastic containers
into which desired vials are retained.
[0584] In some embodiments, kit components are provided in one
and/or more liquid solutions. In some embodiments, liquid solutions
are aqueous solutions, with sterile aqueous solutions being
particularly preferred. In some embodiments, kit components may be
provided as dried powder(s). When reagents and/or components are
provided as dry powders, such powders may be reconstituted by the
addition of suitable volumes of solvent. In some embodiments, it is
envisioned that solvents may also be provided in another container
means. In some embodiments, labeling dyes are provided as dried
powders. In some embodiments, it is contemplated that 10, 20, 30,
40, 50, 60, 70, 80, 90, 100, 120, 120, 130, 140, 150, 160, 170,
180, 190, 200, 300, 400, 500, 600, 700, 800, 900, 1000 micrograms
or at least or at most those amounts of dried dye are provided in
kits of the disclosure. In such embodiments, dye may then be
resuspended in any suitable solvent, such as DMSO.
[0585] In some embodiments, kits may include instructions for
employing kit components as well the use of any other reagent not
included in the kit. Instructions may include variations that may
be implemented.
Devices
[0586] In some embodiments, AAV particles of the present disclosure
may be combined with, coated onto or embedded in a device. Devices
may include, but are not limited to stents, pumps, and/or other
implantable therapeutic device. Additionally AAV particles may be
delivered to a subject while the subject is using a compression
device such as, but not limited to, a compression device to reduce
the chances of deep vein thrombosis (DVT) in a subject.
[0587] The present disclosure provides for devices which may
incorporate AAV particles. These devices contain in a stable
formulation the AAV particles which may be immediately delivered to
a subject in need thereof, such as a human patient.
[0588] Devices for administration may be employed to deliver the
AAV particles of the present disclosure according to single, multi-
or split-dosing regimens taught herein.
[0589] Method and devices known in the art for multi-administration
to cells, organs and tissues are contemplated for use in
conjunction with the methods and compositions disclosed herein as
embodiments of the present disclosure. These include, for example,
those methods and devices having multiple needles, hybrid devices
employing for example lumens or catheters as well as devices
utilizing heat, electric current or radiation driven
mechanisms.
[0590] In some embodiments, AAV particles of the present disclosure
may be delivered using a device such as, but not limited to, a
stent, a tube, a catheter, a pipe, a straw, needle and/or a duct.
Methods of using these devices are described herein and are known
in the art.
[0591] In one embodiment, the AAV particles of the present
disclosure may be administered to a subject using delivery systems
which integrate image guided therapy and integrate imaging such as,
but not limited to, laser, MRgFUS, endoscopic and robotic surgery
devices.
[0592] In one embodiment, the AAV particles of the present
disclosure may be administered to a subject using the
CLEARPOINT.RTM. neuro intervention system by MRI Interventions,
Inc. The CLEARPOINT.RTM. neuro intervention system may be used
alone or in combination with any of the other administration
methods and devices described herein. The CLEARPOINT.RTM. neuro
intervention system helps to provide stereotactic guidance in the
placement and operation of instruments or devices during the
planning and operation of neurological procedures.
[0593] In one embodiment, the AAV particles of the present
disclosure may be administered to a subject using the
NEUROMATE.RTM. stereotactic robot system by Renishaw PLC. The
NEUROMATE.RTM. system may be used alone or in combination with any
of the other administration methods and devices described herein.
As a non-limiting example, the NEUROMATE.RTM. system may be used
with head holders, CT image localizers, frame attachments, remote
controls and software.
[0594] In one embodiment, the AAV particles of the present
disclosure may be administered to a subject using the Elekta
MICRODRIVE.TM. device by Elekta AB. The MICRODRIVE.TM. device may
be used alone or in combination with any of the other
administration methods and devices described herein. As a
non-limiting example, the MICRODRIVE.TM. device may be used to
position electrodes (e.g., for micro electrode recording (MER),
macro stimulation and deep brain stimulation (DBS) electrode
implantation), implantation of catheters, tubes or DBS electrodes
using cross-hair and A-P holders to verify position, biopsies,
injections and aspirations, brain lesioning, endoscope guidance and
GAMMA KNIFE.RTM. radiosurgery.
[0595] In one embodiment, the AAV particles of the present
disclosure may be administered to a subject using the AXIIIS.RTM.
stereotactic miniframe by MONTERIS.RTM. Medical, Inc. The
AXIIIS.RTM. stereotactic miniframe may be used alone or in
combination with any of the other administration methods and
devices described herein. The AXIIIS.RTM. stereotactic miniframe is
a trajectory alignment device which may be used for laser
coagulation, biopsies, catheter placement, electrode implant,
endoscopy, and clot evacuation. The miniframe allows for 360 degree
interface and provides access to multiple intracranial targets with
a simple adjustment. Further, the miniframe is compatible with
MRI.
[0596] In one embodiment, the AAV particles of the present
disclosure may be administered to a subject using the INTEGRA.TM.
CRW.RTM. system by Integra LifeSciences Corporation. The
INTEGRA.TM. CRW.RTM. system may be used alone or in combination
with any of the other administration methods and devices described
herein. The CRW.RTM. system may be used for various applications
such as, but not limited to, stereotactic surgery, microsurgery,
catheterization and biopsy. The CRW.RTM. system is designed to
provide accuracy to those who use the system (e.g., thumb lock
screws, Vernier scaling, double bolt fixation, and a solid
frame).
[0597] In one embodiment, the AAV particles of the present
disclosure may be administered to a subject using the EPOCH.RTM.
solution system by Stereotaxis, Inc. which may include the
NIOBE.RTM. ES magnetic navigation system, the VDRIVE.RTM. robotic
navigation system and/or the ODYSSEY.RTM. information solution (all
by Stereotaxis, Inc.). The EPOCH.RTM. solution system may be used
alone or in combination with any of the other administration
methods and devices described herein. As a non-limiting example,
the NIOBE.RTM. ES magnetic navigation system may be used to
accurately contact a subject. As another non-limiting example the
NIOBE.RTM. ES magnetic system may be used with the VDRIVE.RTM.
robotic navigation system to provide precise movement and
stability.
[0598] In one embodiment, the AAV particles of the present
disclosure may be administered to a subject using a NeuroStation
workstation which uses frameless stereotactic methods to provide
image-guidance for applications such as, but not limited to,
surgical planning, biopsies, craniotomies, endoscopy,
intra-operative ultrasound and radiation therapy.
[0599] In one embodiment, the AAV particles of the present
disclosure may be administered to a subject using a robotic
stereotaxis system such as, but not limited to the device described
in U.S. Pat. No. 5,078,140, the contents of which are herein
incorporated by reference in its entirety. The robotic arm of the
device may be used to precisely orient the surgical tools or other
implements used to conduct a procedure.
[0600] In one embodiment, the AAV particles of the present
disclosure may be administered to a subject using an automatic
delivery system such as, but not limited to the device described in
U.S. Pat. No. 5,865,744, the contents of which are herein
incorporated by reference in its entirety. Based on the images
gathered by the delivery system, the computer adjusts the
administration of the needle to be the appropriate depth for the
particular subject.
[0601] In one embodiment, the AAV particles of the present
disclosure may be administered to a subject who is simultaneously
using during administration, and/or uses for a period of time
before and/or after administration a compression device such as,
but not limited to, a compression device which reduces the chances
of deep vein thrombosis (DVT) in a subject. The compression device
may be used for at least 5 minutes, 15 minutes, 30 minutes, 45
minutes, 1 hour, 2 hours, 3 hours, 4 hours, 5 hours, 6 hours, 7
hours, 8 hours, or more than 8 hours before a subject is
administered the AAV particles. The compression device may be used
for at least 5 minutes, 15 minutes, 30 minutes, 45 minutes, 1 hour,
2 hours, 3 hours, 4 hours, 5 hours, 6 hours, 7 hours, 8 hours, 9
hours, 10 hours, 11 hours, 12 hours, 13 hours, 14 hours, 15 hours,
16 hours, 17 hours, 18 hours, 19 hours, 20 hours, 21 hours, 22
hours, 23 hours, 1 day, 2 days, 3 days, 4 days, 5 days, 6 days, 1
week, 2 weeks, 3 weeks or a month after the AAV particles are
administered. As a non-limiting example, the compression device is
used simultaneously during the procedure of the delivery of the AAV
particles. As another non-limiting example, the compression device
is used before the administration of the AAV particles. As another
non-limiting example, the compression device is used after
administration of the AAV particles. As another non-limiting
example, the compression device is used before, during and after
administration of the AAV particles.
[0602] Non-limiting examples, of compression devices include
ActiveCare+S.F.T. intermittent compression device, ActiveCare+S.F.T
pneumatic compression device, DVTlite's Venowave, KCI system
compression pump, Aircast VenaFlow system, SCD Express Compression
System or Bio Compression Systems, Inc. pneumatic compression
therapy equipment (e.g., the pump may be selected from Model
SC-2004, Model SC-2004-FC, Model SC-3004, Model SC-3004-FC, Model
SC-2008, Model SC-2008-DL, Model SC-3008-T, the BioCryo system,
Model IC-BAP-DL or multi-flo DVT combo IC_1545-DL and the garment
used with the pump may be a 4 chamber, 8 chamber, BioCryo,
Multi-Flo or BioArterial garment).
[0603] In one embodiment, the AAV particles may delivered to a
subject using a device to deliver the AAV particles and a head
fixation assembly. The head fixation assembly may be, but is not
limited to, any of the head fixation assemblies sold by MRI
interventions. As a non-limiting example, the head fixation
assembly may be any of the assemblies described in U.S. Pat. Nos.
8,099,150, 8,548,569 and 9,031,636 and International Patent
Publication Nos. WO201108495 and WO2014014585, the contents of each
of which are incorporated by reference in their entireties. A head
fixation assembly may be used in combination with an MRI compatible
drill such as, but not limited to, the MRI compatible drills
described in International Patent Publication No. WO2013181008 and
US Patent Publication No. US20130325012, the contents of which are
herein incorporated by reference in its entirety.
[0604] In one embodiment, the AAV particles may be delivered using
a method, system and/or computer program for positioning apparatus
to a target point on a subject to deliver the AAV particles. As a
non-limiting example, the method, system and/or computer program
may be the methods, systems and/or computer programs described in
U.S. Pat. No. 8,340,743, the contents of which are herein
incorporated by reference in its entirety. The method may include:
determining a target point in the body and a reference point,
wherein the target point and the reference point define a planned
trajectory line (PTL) extending through each; determining a
visualization plane, wherein the PTL intersects the visualization
plane at a sighting point; mounting the guide device relative to
the body to move with respect to the PTL, wherein the guide device
does not intersect the visualization plane; determining a point of
intersection (GPP) between the guide axis and the visualization
plane; and aligning the GPP with the sighting point in the
visualization plane. In one embodiment, the AAV particles may be
delivered to a subject using a convention-enhanced delivery device.
Non-limiting examples of targeted delivery of drugs using
convection are described in US Patent Publication Nos.
US20100217228, US20130035574 and US20130035660 and International
Patent Publication No. WO2013019830 and WO2008144585, the contents
of each of which are herein incorporated by reference in their
entireties.
[0605] In one embodiment, a subject may be imaged prior to, during
and/or after delivery of the AAV particles. The imaging method may
be a method known in the art and/or described herein, such as but
not limited to, magnetic resonance imaging (MRI). As a non-limiting
example, imaging may be used to assess therapeutic effect. As
another non-limiting example, imaging may be used for assisted
delivery of AAV particles.
[0606] In one embodiment, the AAV particles may be delivered using
an MRI-guided device. Non-limiting examples of MRI-guided devices
are described in U.S. Pat. Nos. 9,055,884, 9,042,958, 8,886,288,
8,768,433, 8,396,532, 8,369,930, 8,374,677 and 8,175,677 and US
Patent Application No. US20140024927 the contents of each of which
are herein incorporated by reference in their entireties. As a
non-limiting example, the MRI-guided device may be able to provide
data in real time such as those described in U.S. Pat. Nos.
8,886,288 and 8,768,433, the contents of each of which is herein
incorporated by reference in its entirety. As another non-limiting
example, the MRI-guided device or system may be used with a
targeting cannula such as the systems described in U.S. Pat. Nos.
8,175,677 and 8,374,677, the contents of each of which are herein
incorporated by reference in their entireties. As yet another
non-limiting example, the MRI-guided device includes a trajectory
guide frame for guiding an interventional device as described, for
example, in U.S. Pat. No. 9,055,884 and US Patent Application No.
US20140024927, the contents of each of which are herein
incorporated by reference in their entireties.
[0607] In one embodiment the AAV particles may be delivered using
an MRI-compatible tip assembly. Non-limiting examples of
MRI-compatible tip assemblies are described in US Patent
Publication No. US20140275980, the contents of which is herein
incorporated by reference in its entirety.
[0608] In one embodiment, the AAV particles may be delivered using
a cannula which is MRI-compatible. Non-limiting examples of
MRI-compatible cannulas include those taught in International
Patent Publication No. WO2011130107, the contents of which are
herein incorporated by reference in its entirety.
[0609] In one embodiment, the AAV particles may be delivered using
a catheter which is MRI-compatible. Non-limiting examples of
MRI-compatible catheters include those taught in International
Patent Publication No. WO2012116265, U.S. Pat. No. 8,825,133 and US
Patent Publication No. US20140024909, the contents of each of which
are herein incorporated by reference in their entireties.
[0610] In one embodiment, the AAV particles may be delivered using
a device with an elongated tubular body and a diaphragm as
described in US Patent Publication Nos. US20140276582 and
US20140276614, the contents of each of which are herein
incorporated by reference in their entireties.
[0611] In one embodiment, the AAV particles may be delivered using
an MRI compatible localization and/or guidance system such as, but
not limited to, those described in US Patent Publication Nos.
US20150223905 and US20150230871, the contents of each of which are
herein incorporated by reference in their entireties. As a
non-limiting example, the MRI compatible localization and/or
guidance systems may comprise a mount adapted for fixation to a
patient, a targeting cannula with a lumen configured to attach to
the mount so as to be able to controllably translate in at least
three dimensions, and an elongate probe configured to snugly
advance via slide and retract in the targeting cannula lumen, the
elongate probe comprising at least one of a stimulation or
recording electrode.
[0612] In one embodiment, the AAV particles may be delivered to a
subject using a trajectory frame as described in US Patent
Publication Nos. US20150031982 and US20140066750 and International
Patent Publication Nos. WO2015057807 and WO2014039481, the contents
of each of which are herein incorporated by reference in their
entireties.
[0613] In one embodiment, the AAV particles may be delivered to a
subject using a gene gun.
Definitions
[0614] At various places in the present specification, substituents
of compounds of the present disclosure are disclosed in groups or
in ranges. It is specifically intended that the present disclosure
include each and every individual subcombination of the members of
such groups and ranges.
[0615] About: As used herein, the term "about" means+/-10% of the
recited value.
[0616] Activity: As used herein, the term "activity" refers to the
condition in which things are happening or being done. Compositions
described herein may have activity and this activity may involve
one or more biological events.
[0617] Adeno-associated virus: The term "adeno-associated virus" or
"AAV" as used herein refers to members of the dependovirus genus
comprising any particle, sequence, gene, protein, or component
derived therefrom. The term "AAV particle" as used herein comprises
a capsid and a polynucleotide. The AAV particle may be derived from
any serotype, described herein or known in the art, including
combinations of serotypes (i.e., "pseudotyped" AAV) or from various
genomes (e.g., single stranded or self-complementary). In addition,
the AAV particle may be replication defective and/or targeted.
[0618] Administered in combination: As used herein, the term
"administered in combination" or "combined administration" means
that two or more agents (e.g., AAV) are administered to a subject
at the same time or within an interval such that there may be an
overlap of an effect of each agent on the patient and/or the
subject is at some point in time simultaneously exposed to both. In
some embodiments, they are administered within about 60, 30, 15,
10, 5, or 1 minutes of one another or within about 24 hours, 12
hours, 6 hours, 3 hours of at least one dose of one or more other
agents. In some embodiments, administration occurs in overlapping
dosage regimens. As used herein, the term "dosage regimen" refers
to a plurality of doses spaced apart in time. Such doses may occur
at regular intervals or may include one or more hiatus in
administration. In some embodiments, the administrations of the
agents are spaced sufficiently closely together such that a
combinatorial (e.g., a synergistic) effect is achieved.
[0619] Amelioration: As used herein, the term "amelioration" or
"ameliorating" refers to a lessening of severity of at least one
indicator of a condition or disease. For example, in the context of
neurodegeneration disorder, amelioration includes the reduction of
neuron loss.
[0620] Animal: As used herein, the term "animal" refers to any
member of the animal kingdom. In some embodiments, "animal" refers
to humans at any stage of development. In some embodiments,
"animal" refers to non-human animals at any stage of development.
In certain embodiments, the non-human animal is a mammal (e.g., a
rodent, a mouse, a rat, a rabbit, a monkey, a dog, a cat, a sheep,
cattle, a primate, or a pig). In some embodiments, animals include,
but are not limited to, mammals, birds, reptiles, amphibians, fish,
and worms. In some embodiments, the animal is a transgenic animal,
genetically-engineered animal, or a clone.
[0621] Antisense strand: As used herein, the term "the antisense
strand" or "the first strand" or "the guide strand" of a siRNA
molecule refers to a strand that is substantially complementary to
a section of about 10-50 nucleotides, e.g., about 15-30, 16-25,
18-23 or 19-22 nucleotides of the mRNA of the gene targeted for
silencing. The antisense strand or first strand has sequence
sufficiently complementary to the desired target mRNA sequence to
direct target-specific silencing, e.g., complementarity sufficient
to trigger the destruction of the desired target mRNA by the RNAi
machinery or process.
[0622] Approximately: As used herein, the term "approximately" or
"about," as applied to one or more values of interest, refers to a
value that is similar to a stated reference value. In certain
embodiments, the term "approximately" or "about" refers to a range
of values that fall within 25%, 20%, 19%, 18%, 17%, 16%, 15%, 14%,
13%, 12%, 11%, 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2%, 1%, or less in
either direction (greater than or less than) of the stated
reference value unless otherwise stated or otherwise evident from
the context (except where such number would exceed 100% of a
possible value).
[0623] Associated with: As used herein, the terms "associated
with," "conjugated," "linked," "attached," and "tethered," when
used with respect to two or more moieties, means that the moieties
are physically associated or connected with one another, either
directly or via one or more additional moieties that serves as a
linking agent, to form a structure that is sufficiently stable so
that the moieties remain physically associated under the conditions
in which the structure is used, e.g., physiological conditions. An
"association" need not be strictly through direct covalent chemical
bonding. It may also suggest ionic or hydrogen bonding or a
hybridization based connectivity sufficiently stable such that the
"associated" entities remain physically associated.
[0624] Bifunctional: As used herein, the term "bifunctional" refers
to any substance, molecule or moiety which is capable of or
maintains at least two functions. The functions may affect the same
outcome or a different outcome. The structure that produces the
function may be the same or different.
[0625] Biologically active: As used herein, the phrase
"biologically active" refers to a characteristic of any substance
(e.g., AAV) that has activity in a biological system and/or
organism. For instance, a substance that, when administered to an
organism, has a biological effect on that organism, is considered
to be biologically active. In particular embodiments, a
polynucleotide of the present disclosure may be considered
biologically active if even a portion of the polynucleotides is
biologically active or mimics an activity considered biologically
relevant.
[0626] Biological system: As used herein, the term "biological
system" refers to a group of organs, tissues, cells, intracellular
components, proteins, nucleic acids, molecules (including, but not
limited to biomolecules) that function together to perform a
certain biological task within cellular membranes, cellular
compartments, cells, tissues, organs, organ systems, multicellular
organisms, or any biological entity. In some embodiments,
biological systems are cell signaling pathways comprising
intracellular and/or extracellular cell signaling biomolecules. In
some embodiments, biological systems comprise growth factor
signaling events within the extracellular/cellular matrix and/or
cellular niches.
[0627] Biomolecule: As used herein, the term "biomolecule" is any
natural molecule which is amino acid-based, nucleic acid-based,
carbohydrate-based or lipid-based, and the like.
[0628] Complementary and substantially complementary: As used
herein, the term "complementary" refers to the ability of
polynucleotides to form base pairs with one another. Base pairs are
typically formed by hydrogen bonds between nucleotide units in
antiparallel polynucleotide strands. Complementary polynucleotide
strands can form base pairs in the Watson-Crick manner (e.g., A to
T, A to U, C to G), or in any other manner that allows for the
formation of duplexes. As persons skilled in the art are aware,
when using RNA as opposed to DNA, uracil rather than thymine is the
base that is considered to be complementary to adenosine. However,
when a U is denoted in the context of the present disclosure, the
ability to substitute a T is implied, unless otherwise stated.
Perfect complementarity or 100% complementarity refers to the
situation in which each nucleotide unit of one polynucleotide
strand can form a hydrogen bond with a nucleotide unit of a second
polynucleotide strand. Less than perfect complementarity refers to
the situation in which some, but not all, nucleotide units of two
strands can form hydrogen bonds with each other. For example, for
two 20-mers, if only two base pairs on each strand can form
hydrogen bonds with each other, the polynucleotide strands exhibit
10% complementarity. In the same example, if 18 base pairs on each
strand can form hydrogen bonds with each other, the polynucleotide
strands exhibit 90% complementarity. As used herein, the term
"substantially complementary" means that the siRNA has a sequence
(e.g., in the antisense strand) which is sufficient to bind the
desired target mRNA, and to trigger the RNA silencing of the target
mRNA.
[0629] Compound: As used herein, the term "compound," refers to a
distinct chemical entity. In some embodiments, a particular
compound may exist in one or more isomeric or isotopic forms
(including, but not limited to stereoisomers, geometric isomers and
isotopes). In some embodiments, a compound is provided or utilized
in only a single such form. In some embodiments, a compound is
provided or utilized as a mixture of two or more such forms
(including, but not limited to a racemic mixture of stereoisomers).
Those of skill in the art appreciate that some compounds exist in
different such forms, show different properties and/or activities
(including, but not limited to biological activities). In such
cases it is within the ordinary skill of those in the art to select
or avoid particular forms of the compound for use in accordance
with the present disclosure. For example, compounds that contain
asymmetrically substituted carbon atoms can be isolated in
optically active or racemic forms. Methods on how to prepare
optically active forms from optically active starting materials are
known in the art, such as by resolution of racemic mixtures or by
stereoselective synthesis. Many geometric isomers of olefins,
C.dbd.N double bonds, and the like can also be present in the
compounds described herein, and all such stable isomers are
contemplated in the present disclosure. Cis and trans geometric
isomers of the compounds of the present disclosure are described
and may be isolated as a mixture of isomers or as separated
isomeric forms.
[0630] Compounds of the present disclosure also include tautomeric
forms. Tautomeric forms result from the swapping of a single bond
with an adjacent double bond and the concomitant migration of a
proton. Tautomeric forms include prototropic tautomers which are
isomeric protonation states having the same empirical formula and
total charge.
[0631] Compounds of the present disclosure also include all of the
isotopes of the atoms occurring in the intermediate or final
compounds. "Isotopes" refers to atoms having the same atomic number
but different mass numbers resulting from a different number of
neutrons in the nuclei. For example, isotopes of hydrogen include
tritium and deuterium.
[0632] The compounds and salts of the present disclosure can be
prepared in combination with solvent or water molecules to form
solvates and hydrates by routine methods.
[0633] Conserved: As used herein, the term "conserved" refers to
nucleotides or amino acid residues of a polynucleotide sequence or
polypeptide sequence, respectively, that are those that occur
unaltered in the same position of two or more sequences being
compared. Nucleotides or amino acids that are relatively conserved
are those that are conserved amongst more related sequences than
nucleotides or amino acids appearing elsewhere in the
sequences.
[0634] In some embodiments, two or more sequences are said to be
"completely conserved" if they are 100% identical to one another.
In some embodiments, two or more sequences are said to be "highly
conserved" if they are at least 70% identical, at least 80%
identical, at least 90% identical, or at least 95% identical to one
another. In some embodiments, two or more sequences are said to be
"highly conserved" if they are about 70% identical, about 80%
identical, about 90% identical, about 95%, about 98%, or about 99%
identical to one another. In some embodiments, two or more
sequences are said to be "conserved" if they are at least 30%
identical, at least 40% identical, at least 50% identical, at least
60% identical, at least 70% identical, at least 80% identical, at
least 90% identical, or at least 95% identical to one another. In
some embodiments, two or more sequences are said to be "conserved"
if they are about 30% identical, about 40% identical, about 50%
identical, about 60% identical, about 70% identical, about 80%
identical, about 90% identical, about 95% identical, about 98%
identical, or about 99% identical to one another. Conservation of
sequence may apply to the entire length of an oligonucleotide, a
polynucleotide or polypeptide or may apply to a portion, region or
feature thereof.
[0635] In one embodiment, conserved sequences are not contiguous.
Those skilled in the art are able to appreciate how to achieve
alignment when gaps in contiguous alignment are present between
sequences, and to align corresponding residues not withstanding
insertions or deletions present.
[0636] In one embodiment, conserved sequences are not contiguous.
Those skilled in the art are able to appreciate how to achieve
alignment when gaps in contiguous alignment are present between
sequences, and to align corresponding residues not withstanding
insertions or deletions present.
[0637] Delivery: As used herein, "delivery" refers to the act or
manner of delivering a compound such as a parvovirus, e.g. an AAV
and/or AAV compound, substance, entity, moiety, cargo or payload to
a target. Such target may be a cell, tissue, organ, organism, or
system (whether biological or production).
[0638] Delivery Agent: As used herein, "delivery agent" refers to
any agent or substance which facilitates, at least in part, the in
vivo and/or in vitro delivery of a polynucleotide and/or one or
more substances (including, but not limited to a compounds and/or
compositions of the present disclosure, e.g., viral particles or
expression vectors) to targeted cells.
[0639] Destabilized: As used herein, the term "destable,"
"destabilize," or "destabilizing region" means a region or molecule
that is less stable than a starting, reference, wild-type or native
form of the same region or molecule.
[0640] Detectable label: As used herein, "detectable label" refers
to one or more markers, signals, or moieties which are attached,
incorporated or associated with another entity that is readily
detected by methods known in the art including radiography,
fluorescence, chemiluminescence, enzymatic activity, absorbance
immunological detection, and the like. Detectable labels may
include radioisotopes, fluorophores, chromophores, enzymes, dyes,
metal ions, ligands such as biotin, avidin, streptavidin and
haptens, quantum dots, and the like. Detectable labels may be
located at any position in the entity with which they are attached,
incorporated or associated. For example, when attached,
incorporated in or associated with a peptide or protein, they may
be within the amino acids, the peptides, or proteins, or located at
the N- or C-termini.
[0641] Dosing regimen: As used herein, a "dosing regimen" is a
schedule of administration or physician determined regimen of
treatment, prophylaxis, or palliative care.
[0642] Effective Amount: As used herein, the term "effective
amount" of an agent is that amount sufficient to effect beneficial
or desired results, for example, upon single or multiple dose
administration to a subject cell, in curing, alleviating, relieving
or improving one or more symptoms of a disorder, clinical results,
and, as such, an "effective amount" depends upon the context in
which it is being applied. For example, in the context of
administering an agent that treats Parkinson's Disease, an
effective amount of an agent is, for example, an amount sufficient
to achieve treatment, as defined herein, of Parkinson's Disease, as
compared to the response obtained without administration of the
agent.
[0643] Encapsulate: As used herein, the term "encapsulate" means to
enclose, surround or encase.
[0644] Engineered: As used herein, embodiments are "engineered"
when they are designed to have a feature or property, whether
structural or chemical, that varies from a starting point,
wild-type or native molecule. Thus, engineered agents or entities
are those whose design and/or production include an act of the hand
of man.
[0645] Epitope: As used herein, an "epitope" refers to a surface or
region on a molecule that is capable of interacting with a
biomolecule. For example a protein may contain one or more amino
acids, e.g., an epitope, which interacts with an antibody, e.g., a
biomolecule. In some embodiments, when referring to a protein or
protein module, an epitope may comprise a linear stretch of amino
acids or a three dimensional structure formed by folded amino acid
chains.
[0646] Expression: As used herein, "expression" of a nucleic acid
sequence refers to one or more of the following events: (1)
production of an RNA template from a DNA sequence (e.g., by
transcription); (2) processing of an RNA transcript (e.g., by
splicing, editing, 5' cap formation, and/or 3' end processing); (3)
translation of an RNA into a polypeptide or protein; (4) folding of
a polypeptide or protein; and (5) post-translational modification
of a polypeptide or protein.
[0647] Feature: As used herein, a "feature" refers to a
characteristic, a property, or a distinctive element.
[0648] Formulation: As used herein, a "formulation" includes at
least one polynucleotide and/or compound and/or composition of the
present disclosure (e.g., a vector, AAV particle, etc.) and a
delivery agent.
[0649] Fragment: A "fragment," as used herein, refers to a
contiguous portion of a whole. For example, fragments of proteins
may comprise polypeptides obtained by digesting full-length protein
isolated from cultured cells. In some embodiments, a fragment of a
protein includes at least 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14,
15, 16, 17, 18, 19, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75,
80, 85, 90, 95, 100, 150, 200, 250 or more amino acids. In some
embodiments, fragments of an antibody include portions of an
antibody subjected to enzymatic digestion or synthesized as
such.
[0650] Functional: As used herein, a "functional" biological
molecule is a biological molecule and/or entity with a structure
and in a form in which it exhibits a property and/or activity by
which it is characterized.
[0651] Gene expression: The term "gene expression" refers to the
process by which a nucleic acid sequence undergoes successful
transcription and in most instances translation to produce a
protein or peptide. For clarity, when reference is made to
measurement of "gene expression", this should be understood to mean
that measurements may be of the nucleic acid product of
transcription, e.g., RNA or mRNA or of the amino acid product of
translation, e.g., polypeptides or peptides. Methods of measuring
the amount or levels of RNA, mRNA, polypeptides and peptides are
well known in the art.
[0652] Homology: As used herein, the term "homology" refers to the
overall relatedness between polymeric molecules, e.g. between
nucleic acid molecules (e.g. DNA molecules and/or RNA molecules)
and/or between polypeptide molecules. In some embodiments,
polymeric molecules are considered to be "homologous" to one
another if their sequences are at least 25%, 30%, 35%, 40%, 45%,
50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 99% identical
or similar. The term "homologous" necessarily refers to a
comparison between at least two sequences (polynucleotide or
polypeptide sequences). In accordance with the disclosure, two
polynucleotide sequences are considered to be homologous if the
polypeptides they encode are at least about 50%, 60%, 70%, 80%,
90%, 95%, or even 99% for at least one stretch of at least about 20
amino acids. In some embodiments, homologous polynucleotide
sequences are characterized by the ability to encode a stretch of
at least 4-5 uniquely specified amino acids. For polynucleotide
sequences less than 60 nucleotides in length, homology is typically
determined by the ability to encode a stretch of at least 4-5
uniquely specified amino acids. In accordance with the disclosure,
two protein sequences are considered to be homologous if the
proteins are at least about 50%, 60%, 70%, 80%, or 90% identical
for at least one stretch of at least about 20 amino acids. In many
embodiments, homologous protein may show a large overall degree of
homology and a high degree of homology over at least one short
stretch of at least 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15,
16, 17, 18, 19, 20, 25, 30, 35, 40, 45, 50 or more amino acids. In
many embodiments, homologous proteins share one or more
characteristic sequence elements. As used herein, the term
"characteristic sequence element" refers to a motif present in
related proteins. In some embodiments, the presence of such motifs
correlates with a particular activity (such as biological
activity).
[0653] Identity: As used herein, the term "identity" refers to the
overall relatedness between polymeric molecules, e.g., between
oligonucleotide and/or polynucleotide molecules (e.g. DNA molecules
and/or RNA molecules) and/or between polypeptide molecules.
Calculation of the percent identity of two polynucleotide
sequences, for example, may be performed by aligning the two
sequences for optimal comparison purposes (e.g., gaps can be
introduced in one or both of a first and a second nucleic acid
sequences for optimal alignment and non-identical sequences can be
disregarded for comparison purposes). In certain embodiments, the
length of a sequence aligned for comparison purposes is at least
30%, at least 40%, at least 50%, at least 60%, at least 70%, at
least 80%, at least 90%, at least 95%, or 100% of the length of the
reference sequence. The nucleotides at corresponding nucleotide
positions are then compared. When a position in the first sequence
is occupied by the same nucleotide as the corresponding position in
the second sequence, then the molecules are identical at that
position. The percent identity between the two sequences is a
function of the number of identical positions shared by the
sequences, taking into account the number of gaps, and the length
of each gap, which needs to be introduced for optimal alignment of
the two sequences. The comparison of sequences and determination of
percent identity between two sequences can be accomplished using a
mathematical algorithm. For example, the percent identity between
two nucleotide sequences can be determined using methods such as
those described in Computational Molecular Biology, Lesk, A. M.,
ed., Oxford University Press, New York, 1988; Biocomputing:
Informatics and Genome Projects, Smith, D. W., ed., Academic Press,
New York, 1993; Sequence Analysis in Molecular Biology, von Heinje,
G., Academic Press, 1987; Computer Analysis of Sequence Data, Part
I, Griffin, A. M., and Griffin, H. G., eds., Humana Press, New
Jersey, 1994; and Sequence Analysis Primer, Gribskov, M. and
Devereux, J., eds., M Stockton Press, New York, 1991; each of which
is incorporated herein by reference in its entirety. For example,
the percent identity between two nucleotide sequences can be
determined, for example using the algorithm of Meyers and Miller
(CABIOS, 1989, 4:11-17), which has been incorporated into the ALIGN
program (version 2.0) using a PAM120 weight residue table, a gap
length penalty of 12 and a gap penalty of 4. The percent identity
between two nucleotide sequences can, alternatively, be determined
using the GAP program in the GCG software package using an
NWSgapdna.CMP matrix. Methods commonly employed to determine
percent identity between sequences include, but are not limited to
those disclosed in Carillo, H., and Lipman, D., SIAM J Applied
Math., 48:1073 (1988); incorporated herein by reference in its
entirety. Techniques for determining identity are codified in
publicly available computer programs. Exemplary computer software
to determine homology between two sequences include, but are not
limited to, GCG program package, Devereux, J., et al., Nucleic
Acids Research, 12(1), 387 (1984)), BLASTP, BLASTN, and FASTA
Altschul, S. F. et al., J. Molec. Biol., 215, 403 (1990)).
[0654] Inhibit expression of a gene: As used herein, the phrase
"inhibit expression of a gene" means to cause a reduction in the
amount of an expression product of the gene. The expression product
may be RNA transcribed from the gene (e.g. mRNA) or a polypeptide
translated from mRNA transcribed from the gene. Typically a
reduction in the level of mRNA results in a reduction in the level
of a polypeptide translated therefrom. The level of expression may
be determined using standard techniques for measuring mRNA or
protein.
[0655] In vitro: As used herein, the term "in vitro" refers to
events that occur in an artificial environment, e.g., in a test
tube or reaction vessel, in cell culture, in a Petri dish, etc.,
rather than within an organism (e.g., animal, plant, or
microbe).
[0656] In vivo: As used herein, the term "in vivo" refers to events
that occur within an organism (e.g., animal, plant, or microbe or
cell or tissue thereof).
[0657] Isolated: As used herein, the term "isolated" is synonymous
with "separated", but carries with it the inference separation was
carried out by the hand of man. In one embodiment, an isolated
substance or entity is one that has been separated from at least
some of the components with which it was previously associated
(whether in nature or in an experimental setting). Isolated
substances may have varying levels of purity in reference to the
substances from which they have been associated. Isolated
substances and/or entities may be separated from at least about
10%, about 20%, about 30%, about 40%, about 50%, about 60%, about
70%, about 80%, about 90%, or more of the other components with
which they were initially associated. In some embodiments, isolated
agents are more than about 80%, about 85%, about 90%, about 91%,
about 92%, about 93%, about 94%, about 95%, about 96%, about 97%,
about 98%, about 99%, or more than about 99% pure. As used herein,
a substance is "pure" if it is substantially free of other
components.
[0658] Substantially isolated: By "substantially isolated" is meant
that the compound is substantially separated from the environment
in which it was formed or detected. Partial separation can include,
for example, a composition enriched in the compound of the present
disclosure. Substantial separation can include compositions
containing at least about 50%, at least about 60%, at least about
70%, at least about 80%, at least about 90%, at least about 95%, at
least about 97%, or at least about 99% by weight of the compound of
the present disclosure, or salt thereof. Methods for isolating
compounds and their salts are routine in the art. In some
embodiments, isolation of a substance or entity includes disruption
of chemical associations and/or bonds. In some embodiments,
isolation includes only the separation from components with which
the isolated substance or entity was previously combined and does
not include such disruption.
[0659] Modified: As used herein, the term "modified" refers to a
changed state or structure of a molecule or entity of the
disclosure as compared with a parent or reference molecule or
entity. Molecules may be modified in many ways including
chemically, structurally, and functionally. In some embodiments,
compounds and/or compositions of the present disclosure are
modified by the introduction of non-natural amino acids, or
non-natural nucleotides.
[0660] Mutation: As used herein, the term "mutation" refers to a
change and/or alteration. In some embodiments, mutations may be
changes and/or alterations to proteins (including peptides and
polypeptides) and/or nucleic acids (including polynucleic acids).
In some embodiments, mutations comprise changes and/or alterations
to a protein and/or nucleic acid sequence. Such changes and/or
alterations may comprise the addition, substitution and or deletion
of one or more amino acids (in the case of proteins and/or
peptides) and/or nucleotides (in the case of nucleic acids and or
polynucleic acids). In embodiments wherein mutations comprise the
addition and/or substitution of amino acids and/or nucleotides,
such additions and/or substitutions may comprise 1 or more amino
acid and/or nucleotide residues and may include modified amino
acids and/or nucleotides.
[0661] Naturally occurring: As used herein, "naturally occurring"
means existing in nature without artificial aid or involvement of
the hand of man
[0662] Non-human vertebrate: As used herein, a "non-human
vertebrate" includes all vertebrates except Homo sapiens, including
wild and domesticated species. Examples of non-human vertebrates
include, but are not limited to, mammals, such as alpaca, banteng,
bison, camel, cat, cattle, deer, dog, donkey, gayal, goat, guinea
pig, horse, llama, mule, pig, rabbit, reindeer, sheep water
buffalo, and yak.
[0663] Nucleic acid: As used herein, the term "nucleic acid",
"polynucleotide" and `oligonucleotide" refer to any nucleic acid
polymers composed of either polydeoxyribonucleotides (containing
2-deoxy-D-ribose), or polyribonucleotides (containing D-ribose), or
any other type of polynucleotide which is an N glycoside of a
purine or pyrimidine base, or modified purine or pyrimidine bases.
There is no intended distinction in length between the term
"nucleic acid", "polynucleotide" and "oligonucleotide", and these
terms will be used interchangeably. These terms refer only to the
primary structure of the molecule. Thus, these terms include
double- and single-stranded DNA, as well as double- and single
stranded RNA.
[0664] Off-target: As used herein, "off target" refers to any
unintended effect on any one or more target, gene and/or cellular
transcript.
[0665] Open reading frame: As used herein, "open reading frame" or
"ORF" refers to a sequence which does not contain a stop codon in a
given reading frame.
[0666] Operably linked. As used herein, the phrase "operably
linked" refers to a functional connection between two or more
molecules, constructs, transcripts, entities, moieties or the
like.
[0667] Particle: As used herein, a "particle" is a virus comprised
of at least two components, a protein capsid and a polynucleotide
sequence enclosed within the capsid.
[0668] Patient. As used herein, "patient" refers to a subject who
may seek or be in need of treatment, requires treatment, is
receiving treatment, will receive treatment, or a subject who is
under care by a trained professional for a particular disease or
condition, such as for example Parkinson's Disease.
[0669] Payload: As used herein, "payload" refers to one or more
polynucleotides or polynucleotide regions encoded by or within a
viral genome or an expression product of such polynucleotide or
polynucleotide region, e.g., a transgene, a polynucleotide encoding
a polypeptide or multi-polypeptide or a modulatory nucleic acid or
regulatory nucleic acid.
[0670] Payload construct: As used herein, "payload construct" is
one or more polynucleotide regions encoding or comprising a payload
that is flanked on one or both sides by an inverted terminal repeat
(ITR) sequence. The payload construct is a template that is
replicated in a viral production cell to produce a viral
genome.
[0671] Payload construct vector: As used herein, "payload construct
vector" is a vector encoding or comprising a payload construct, and
regulatory regions for replication and expression in bacterial
cells.
[0672] Payload construct expression vector: As used herein, a
"payload construct expression vector" is a vector encoding or
comprising a payload construct and which further comprises one or
more polynucleotide regions encoding or comprising components for
viral expression in a viral replication cell.
[0673] Peptide: As used herein, "peptide" is less than or equal to
50 amino acids long, e.g., about 5, 10, 15, 20, 25, 30, 35, 40, 45,
or 50 amino acids long.
[0674] Pharmaceutically acceptable: The phrase "pharmaceutically
acceptable" is employed herein to refer to those compounds,
materials, compositions, and/or dosage forms which are, within the
scope of sound medical judgment, suitable for use in contact with
the tissues of human beings and animals without excessive toxicity,
irritation, allergic response, or other problem or complication,
commensurate with a reasonable benefit/risk ratio.
[0675] Pharmaceutically acceptable excipients: The phrase
"pharmaceutically acceptable excipient," as used herein, refers to
any ingredient other than the compounds and/or active agents (e.g.
as described herein) present in pharmaceutical compositions and
having the properties of being substantially nontoxic and
non-inflammatory in a subject such as a patient. In some
embodiments, pharmaceutically acceptable excipients are vehicles
capable of suspending and/or dissolving active agents. Excipients
may include, for example: antiadherents, antioxidants, binders,
coatings, compression aids, disintegrants, dyes (colors),
emollients, emulsifiers, fillers (diluents), film formers or
coatings, flavors, fragrances, glidants (flow enhancers),
lubricants, preservatives, printing inks, sorbents, suspension or
dispersing agents, sweeteners, and waters of hydration. Exemplary
excipients include, but are not limited to: butylated
hydroxytoluene (BHT), calcium carbonate, calcium phosphate
(dibasic), calcium stearate, croscarmellose, crosslinked polyvinyl
pyrrolidone, citric acid, crospovidone, cysteine, ethylcellulose,
gelatin, hydroxypropyl cellulose, hydroxypropyl methylcellulose,
lactose, magnesium stearate, maltitol, mannitol, methionine,
methylcellulose, methyl paraben, microcrystalline cellulose,
polyethylene glycol, polyvinyl pyrrolidone, povidone,
pregelatinized starch, propyl paraben, retinyl palmitate, shellac,
silicon dioxide, sodium carboxymethyl cellulose, sodium citrate,
sodium starch glycolate, sorbitol, starch (corn), stearic acid,
sucrose, talc, titanium dioxide, vitamin A, vitamin E, vitamin C,
and xylitol.
[0676] Pharmaceutically acceptable salts: Pharmaceutically
acceptable salts of the compounds described herein. As used herein,
"pharmaceutically acceptable salts" refers to derivatives or forms
of the disclosed compounds wherein the parent compound is modified
by converting an existing acid or base moiety to its salt form
(e.g., as generated by reacting the free base group with a suitable
organic acid). Examples of pharmaceutically acceptable salts
include, but are not limited to, mineral or organic acid salts of
basic residues such as amines; alkali or organic salts of acidic
residues such as carboxylic acids; and the like. Representative
acid addition salts include acetate, acetic acid, adipate,
alginate, ascorbate, aspartate, benzenesulfonate, benzene sulfonic
acid, benzoate, bisulfate, borate, butyrate, camphorate,
camphorsulfonate, citrate, cyclopentanepropionate, digluconate,
dodecylsulfate, ethanesulfonate, fumarate, glucoheptonate,
glycerophosphate, hemisulfate, heptonate, hexanoate, hydrobromide,
hydrochloride, hydroiodide, 2-hydroxy-ethanesulfonate,
lactobionate, lactate, laurate, lauryl sulfate, malate, maleate,
malonate, methanesulfonate, 2-naphthalenesulfonate, nicotinate,
nitrate, oleate, oxalate, palmitate, pamoate, pectinate,
persulfate, 3-phenylpropionate, phosphate, picrate, pivalate,
propionate, stearate, succinate, sulfate, tartrate, thiocyanate,
toluenesulfonate, undecanoate, valerate salts, and the like.
Representative alkali or alkaline earth metal salts include sodium,
lithium, potassium, calcium, magnesium, and the like, as well as
nontoxic ammonium, quaternary ammonium, and amine cations,
including, but not limited to ammonium, tetramethylammonium,
tetraethylammonium, methylamine, dimethylamine, trimethylamine,
triethylamine, ethylamine, and the like. The pharmaceutically
acceptable salts of the present disclosure include the conventional
non-toxic salts of the parent compound formed, for example, from
non-toxic inorganic or organic acids. In some embodiments a
pharmaceutically acceptable salt of the present disclosure can be
synthesized salt prepared from the parent compound which contains a
basic or acidic moiety by conventional chemical methods. Generally,
such salts can be prepared by reacting the free acid or base forms
of these compounds with a stoichiometric amount of the appropriate
base or acid in water or in an organic solvent, or in a mixture of
the two; generally, nonaqueous media like ether, ethyl acetate,
ethanol, isopropanol, or acetonitrile are preferred. Lists of
suitable salts are found in Remington's Pharmaceutical Sciences,
17th ed., Mack Publishing Company, Easton, Pa., 1985, p. 1418,
Pharmaceutical Salts: Properties, Selection, and Use, P. H. Stahl
and C. G. Wermuth (eds.), Wiley-VCH, 2008, and Berge et al.,
Journal of Pharmaceutical Science, 66, 1-19 (1977), each of which
is incorporated herein by reference in its entirety.
[0677] Pharmaceutically acceptable solvate: The term
"pharmaceutically acceptable solvate," as used herein, refers to a
crystalline form of a compound of the disclosure wherein molecules
of a suitable solvent are incorporated in the crystal lattice. A
suitable solvent is physiologically tolerable at the dosage
administered. For example, solvates may be prepared by
crystallization, recrystallization, or precipitation from a
solution that includes organic solvents, water, or a mixture
thereof. Examples of suitable solvents are ethanol, water (for
example, mono-, di-, and tri-hydrates), N-methylpyrrolidinone
(NMP), dimethyl sulfoxide (DMSO), N,N'-dimethylformamide (DMF),
N,N'-dimethylacetamide (DMAC), 1,3-dimethyl-2-imidazolidinone
(DMEU), 1,3-dimethyl-3,4,5,6-tetrahydro-2-(1H)-pyrimidinone (DMPU),
acetonitrile (ACN), propylene glycol, ethyl acetate, benzyl
alcohol, 2-pyrrolidone, benzyl benzoate, and the like. When water
is the solvent, the solvate is referred to as a "hydrate." In some
embodiments, the solvent incorporated into a solvate is of a type
or at a level that is physiologically tolerable to an organism to
which the solvate is administered (e.g., in a unit dosage form of a
pharmaceutical composition).
[0678] Pharmacokinetic: As used herein, "pharmacokinetic" refers to
any one or more properties of a molecule or compound as it relates
to the determination of the fate of substances administered to a
living organism. Pharmacokinetics is divided into several areas
including the extent and rate of absorption, distribution,
metabolism and excretion. This is commonly referred to as ADME
where: (A) Absorption is the process of a substance entering the
blood circulation; (D) Distribution is the dispersion or
dissemination of substances throughout the fluids and tissues of
the body; (M) Metabolism (or Biotransformation) is the irreversible
transformation of parent compounds into daughter metabolites; and
(E) Excretion (or Elimination) refers to the elimination of the
substances from the body. In rare cases, some drugs irreversibly
accumulate in body tissue.
[0679] Physicochemical. As used herein, "physicochemical" means of
or relating to a physical and/or chemical property.
[0680] Preventing: As used herein, the term "preventing" refers to
partially or completely delaying onset of an infection, disease,
disorder and/or condition; partially or completely delaying onset
of one or more symptoms, features, or clinical manifestations of a
particular infection, disease, disorder, and/or condition;
partially or completely delaying onset of one or more symptoms,
features, or manifestations of a particular infection, disease,
disorder, and/or condition; partially or completely delaying
progression from an infection, a particular disease, disorder
and/or condition; and/or decreasing the risk of developing
pathology associated with the infection, the disease, disorder,
and/or condition, such as for example Parkinson's Disease.
[0681] Prodrug: The present disclosure also includes prodrugs of
the compounds described herein. As used herein, "prodrugs" refer to
any substance, molecule or entity which is in a form predicate for
that substance, molecule or entity to act as a therapeutic upon
chemical or physical alteration. Prodrugs may by covalently bonded
or sequestered in some way and which release or are converted into
the active drug moiety prior to, upon or after administered to a
mammalian subject. Preparation and use of prodrugs is discussed in
T. Higuchi and V. Stella, "Pro-drugs as Novel Delivery Systems,"
Vol. 14 of the A.C.S. Symposium Series, and in Bioreversible
Carriers in Drug Design, ed. Edward B. Roche, American
Pharmaceutical Association and Pergamon Press, 1987, both of which
are hereby incorporated by reference in their entirety.
[0682] Proliferate: As used herein, the term "proliferate" means to
grow, expand, replicate or increase or cause to grow, expand,
replicate or increase. "Proliferative" means having the ability to
proliferate. "Anti-proliferative" means having properties counter
to or in opposition to proliferative properties.
[0683] Prophylactic: As used herein, "prophylactic" refers to a
therapeutic or course of action used to prevent the spread of
disease.
[0684] Prophylaxis: As used herein, a "prophylaxis" refers to a
measure taken to maintain health and prevent the spread of
disease.
[0685] Protein of interest: As used herein, the terms "proteins of
interest" or "desired proteins" include those provided herein and
fragments, mutants, variants, and alterations thereof.
[0686] Purified: As used herein, "purify," "purified,"
"purification" means to make substantially pure or clear from
unwanted components, material defilement, admixture or
imperfection. "Purified" refers to the state of being pure.
"Purification" refers to the process of making pure.
[0687] Region: As used herein, the term "region" refers to a zone
or general area. In some embodiments, when referring to a protein
or protein module, a region may comprise a linear sequence of amino
acids along the protein or protein module or may comprise a three
dimensional area, an epitope and/or a cluster of epitopes. In some
embodiments, regions comprise terminal regions. As used herein, the
term "terminal region" refers to regions located at the ends or
termini of a given agent. When referring to proteins, terminal
regions may comprise N- and/or C-termini. N-termini refer to the
end of a protein comprising an amino acid with a free amino group.
C-termini refer to the end of a protein comprising an amino acid
with a free carboxyl group. N- and/or C-terminal regions may
therefore comprise the N- and/or C-termini as well as surrounding
amino acids. In some embodiments, N- and/or C-terminal regions
comprise from about 3 amino acid to about 30 amino acids, from
about 5 amino acids to about 40 amino acids, from about 10 amino
acids to about 50 amino acids, from about 20 amino acids to about
100 amino acids and/or at least 100 amino acids. In some
embodiments, N-terminal regions may comprise any length of amino
acids that includes the N-terminus, but does not include the
C-terminus. In some embodiments, C-terminal regions may comprise
any length of amino acids, which include the C-terminus, but do not
comprise the N-terminus.
[0688] In some embodiments, when referring to a polynucleotide, a
region may comprise a linear sequence of nucleic acids along the
polynucleotide or may comprise a three dimensional area, secondary
structure, or tertiary structure. In some embodiments, regions
comprise terminal regions. As used herein, the term "terminal
region" refers to regions located at the ends or termini of a given
agent. When referring to polynucleotides, terminal regions may
comprise 5' and 3' termini. 5' termini refer to the end of a
polynucleotide comprising a nucleic acid with a free phosphate
group. 3' termini refer to the end of a polynucleotide comprising a
nucleic acid with a free hydroxyl group. 5' and 3' regions may
therefore comprise the 5' and 3' termini as well as surrounding
nucleic acids. In some embodiments, 5' and 3' terminal regions
comprise from about 9 nucleic acids to about 90 nucleic acids, from
about 15 nucleic acids to about 120 nucleic acids, from about 30
nucleic acids to about 150 nucleic acids, from about 60 nucleic
acids to about 300 nucleic acids and/or at least 300 nucleic acids.
In some embodiments, 5' regions may comprise any length of nucleic
acids that includes the 5' terminus, but does not include the 3'
terminus. In some embodiments, 3' regions may comprise any length
of nucleic acids, which include the 3' terminus, but does not
comprise the 5' terminus.
[0689] RNA or RNA molecule: As used herein, the term "RNA" or "RNA
molecule" or "ribonucleic acid molecule" refers to a polymer of
ribonucleotides; the term "DNA" or "DNA molecule" or
"deoxyribonucleic acid molecule" refers to a polymer of
deoxyribonucleotides. DNA and RNA can be synthesized naturally,
e.g., by DNA replication and transcription of DNA, respectively; or
be chemically synthesized. DNA and RNA can be single-stranded
(i.e., ssRNA or ssDNA, respectively) or multi-stranded (e.g.,
double stranded, i.e., dsRNA and dsDNA, respectively). The term
"mRNA" or "messenger RNA", as used herein, refers to a single
stranded RNA that encodes the amino acid sequence of one or more
polypeptide chains.
[0690] RNA interference: As used herein, the term "RNA
interference" or "RNAi" refers to a sequence specific regulatory
mechanism mediated by RNA molecules which results in the inhibition
or interference or "silencing" of the expression of a corresponding
protein-coding gene.
[0691] Sample. As used herein, the term "sample" refers to an
aliquot, subset or portion taken from a source and/or provided for
analysis or processing. In some embodiments, a sample is from a
biological source such as a tissue, cell or component part (e.g. a
body fluid, including but not limited to blood, mucus, lymphatic
fluid, synovial fluid, cerebrospinal fluid, saliva, amniotic fluid,
amniotic cord blood, urine, vaginal fluid and semen). In some
embodiments, a sample may be or comprise a homogenate, lysate or
extract prepared from a whole organism or a subset of its tissues,
cells or component parts, or a fraction or portion thereof,
including but not limited to, for example, plasma, serum, spinal
fluid, lymph fluid, the external sections of the skin, respiratory,
intestinal, and genitourinary tracts, tears, saliva, milk, blood
cells, tumors, or organs. In some embodiments, a sample is or
comprises a medium, such as a nutrient broth or gel, which may
contain cellular components, such as proteins or nucleic acid
molecule. In some embodiments, a "primary" sample is an aliquot of
the source. In some embodiments, a primary sample is subjected to
one or more processing (e.g., separation, purification, etc.) steps
to prepare a sample for analysis or other use.
[0692] Self-complementary viral particle: As used herein, a
"self-complementary viral particle" is a particle comprised of at
least two components, a protein capsid and a polynucleotide
sequence encoding a self-complementary genome enclosed within the
capsid.
[0693] Sense strand: As used herein, the term "the sense strand" or
"the second strand" or "the passenger strand" of a siRNA molecule
refers to a strand that is complementary to the antisense strand or
first strand. The antisense and sense strands of a siRNA molecule
are hybridized to form a duplex structure. As used herein, a "siRNA
duplex" includes a siRNA strand having sufficient complementarity
to a section of about 10-50 nucleotides of the mRNA of the gene
targeted for silencing and a siRNA strand having sufficient
complementarity to form a duplex with the siRNA strand.
[0694] Signal Sequences: As used herein, the phrase "signal
sequences" refers to a sequence which can direct the transport or
localization.
[0695] Single unit dose: As used herein, a "single unit dose" is a
dose of any therapeutic administered in one dose/at one time/single
route/single point of contact, i.e., single administration event.
In some embodiments, a single unit dose is provided as a discrete
dosage form (e.g., a tablet, capsule, patch, loaded syringe, vial,
etc.).
[0696] Similarity: As used herein, the term "similarity" refers to
the overall relatedness between polymeric molecules, e.g. between
polynucleotide molecules (e.g. DNA molecules and/or RNA molecules)
and/or between polypeptide molecules. Calculation of percent
similarity of polymeric molecules to one another can be performed
in the same manner as a calculation of percent identity, except
that calculation of percent similarity takes into account
conservative substitutions as is understood in the art.
[0697] Small/short interfering RNA: As used herein, the term
"small/short interfering RNA" or "siRNA" refers to an RNA molecule
(or RNA analog) comprising between about 5-60 nucleotides (or
nucleotide analogs) which is capable of directing or mediating
RNAi. Preferably, a siRNA molecule comprises between about 15-30
nucleotides or nucleotide analogs, more preferably between about
16-25 nucleotides (or nucleotide analogs), even more preferably
between about 18-23 nucleotides (or nucleotide analogs), and even
more preferably between about 19-22 nucleotides (or nucleotide
analogs) (e.g., 19, 20, 21 or 22 nucleotides or nucleotide
analogs). The term "short" siRNA refers to a siRNA comprising 5-23
nucleotides, preferably 21 nucleotides (or nucleotide analogs), for
example, 19, 20, 21 or 22 nucleotides. The term "long" siRNA refers
to a siRNA comprising 24-60 nucleotides, preferably about 24-25
nucleotides, for example, 23, 24, 25 or 26 nucleotides. Short
siRNAs may, in some instances, include fewer than 19 nucleotides,
e.g., 16, 17 or 18 nucleotides, or as few as 5 nucleotides,
provided that the shorter siRNA retains the ability to mediate
RNAi. Likewise, long siRNAs may, in some instances, include more
than 26 nucleotides, e.g., 27, 28, 29, 30, 35, 40, 45, 50, 55, or
even 60 nucleotides, provided that the longer siRNA retains the
ability to mediate RNAi or translational repression absent further
processing, e.g., enzymatic processing, to a short siRNA. siRNAs
can be single stranded RNA molecules (ss-siRNAs) or double stranded
RNA molecules (ds-siRNAs) comprising a sense strand and an
antisense strand which hybridized to form a duplex structure called
siRNA duplex.
[0698] Split dose: As used herein, a "split dose" is the division
of single unit dose or total daily dose into two or more doses.
[0699] Stable: As used herein "stable" refers to a compound or
entity that is sufficiently robust to survive isolation to a useful
degree of purity from a reaction mixture, and preferably capable of
formulation into an efficacious therapeutic agent.
[0700] Stabilized: As used herein, the term "stabilize",
"stabilized," "stabilized region" means to make or become stable.
In some embodiments, stability is measured relative to an absolute
value. In some embodiments, stability is measured relative to a
reference compound or entity.
[0701] Subject: As used herein, the term "subject" or "patient"
refers to any organism to which a composition in accordance with
the disclosure may be administered, e.g., for experimental,
diagnostic, prophylactic, and/or therapeutic purposes. Typical
subjects include animals (e.g., mammals such as mice, rats,
rabbits, non-human primates, and humans) and/or plants. In some
embodiments, the subject may be an infant, neonate, or a child
under the age of 12 years old. In some embodiments, the subject may
be in utero.
[0702] Substantially: As used herein, the term "substantially"
refers to the qualitative condition of exhibiting total or
near-total extent or degree of a characteristic or property of
interest. One of ordinary skill in the biological arts will
understand that biological and chemical phenomena rarely, if ever,
go to completion and/or proceed to completeness or achieve or avoid
an absolute result. The term "substantially" is therefore used
herein to capture the potential lack of completeness inherent in
many biological and chemical phenomena.
[0703] Substantially equal: As used herein as it relates to time
differences between doses, the term means plus/minus 2%.
[0704] Substantially simultaneously: As used herein and as it
relates to plurality of doses, the term typically means within
about 2 seconds.
[0705] Suffering from: An individual who is "suffering from" a
disease, disorder, and/or condition has been diagnosed with or
displays one or more symptoms of a disease, disorder, and/or
condition such as for example Parkinson's Disease.
[0706] Susceptible to: An individual who is "susceptible to" a
disease, disorder, and/or condition has not been diagnosed with
and/or may not exhibit symptoms of the disease, disorder, and/or
condition but harbors a propensity to develop a disease or its
symptoms. In some embodiments, an individual who is susceptible to
a disease, disorder, and/or condition (for example, cancer) may be
characterized by one or more of the following: (1) a genetic
mutation associated with development of the disease, disorder,
and/or condition; (2) a genetic polymorphism associated with
development of the disease, disorder, and/or condition; (3)
increased and/or decreased expression and/or activity of a protein
and/or nucleic acid associated with the disease, disorder, and/or
condition; (4) habits and/or lifestyles associated with development
of the disease, disorder, and/or condition; (5) a family history of
the disease, disorder, and/or condition; and (6) exposure to and/or
infection with a microbe associated with development of the
disease, disorder, and/or condition. In some embodiments, an
individual who is susceptible to a disease, disorder, and/or
condition will develop the disease, disorder, and/or condition. In
some embodiments, an individual who is susceptible to a disease,
disorder, and/or condition will not develop the disease, disorder,
and/or condition.
[0707] Sustained release: As used herein, the term "sustained
release" refers to a pharmaceutical composition or compound release
profile that conforms to a release rate over a specific period of
time.
[0708] Synthetic: The term "synthetic" means produced, prepared,
and/or manufactured by the hand of man. Synthesis of
polynucleotides or polypeptides or other molecules of the present
disclosure may be chemical or enzymatic.
[0709] Targeting: As used herein, "targeting" means the process of
design and selection of nucleic acid sequence that will hybridize
to a target nucleic acid and induce a desired effect.
[0710] Targeted Cells: As used herein, "targeted cells" refers to
any one or more cells of interest. The cells may be found in vitro,
in vivo, in situ or in the tissue or organ of an organism. The
organism may be an animal, preferably a mammal, more preferably a
human and most preferably a patient.
[0711] Therapeutic Agent: The term "therapeutic agent" refers to
any agent that, when administered to a subject, has a therapeutic,
diagnostic, and/or prophylactic effect and/or elicits a desired
biological and/or pharmacological effect.
[0712] Therapeutically effective amount: As used herein, the term
"therapeutically effective amount" means an amount of an agent to
be delivered (e.g., nucleic acid, drug, therapeutic agent,
diagnostic agent, prophylactic agent, etc.) that is sufficient,
when administered to a subject suffering from or susceptible to an
infection, disease, disorder, and/or condition, to treat, improve
symptoms of, diagnose, prevent, and/or delay the onset of the
infection, disease, disorder, and/or condition such as for example
Parkinson's Disease. In some embodiments, a therapeutically
effective amount is provided in a single dose. In some embodiments,
a therapeutically effective amount is administered in a dosage
regimen comprising a plurality of doses. Those skilled in the art
will appreciate that in some embodiments, a unit dosage form may be
considered to comprise a therapeutically effective amount of a
particular agent or entity if it comprises an amount that is
effective when administered as part of such a dosage regimen.
[0713] Therapeutically effective outcome: As used herein, the term
"therapeutically effective outcome" means an outcome that is
sufficient in a subject suffering from or susceptible to an
infection, disease, disorder, and/or condition, to treat, improve
symptoms of, diagnose, prevent, and/or delay the onset of the
infection, disease, disorder, and/or condition.
[0714] Total daily dose: As used herein, a "total daily dose" is an
amount given or prescribed in 24 hour period. It may be
administered as a single unit dose.
[0715] Transfection: As used herein, the term "transfection" refers
to methods to introduce exogenous nucleic acids into a cell.
Methods of transfection include, but are not limited to, chemical
methods, physical treatments and cationic lipids or mixtures.
[0716] Treating: As used herein, the term "treating" refers to
partially or completely alleviating, ameliorating, improving,
relieving, delaying onset of, inhibiting progression of, reducing
severity of, and/or reducing incidence of one or more symptoms or
features of a particular infection, disease, disorder, and/or
condition. For example, "treating" cancer may refer to inhibiting
survival, growth, and/or spread of a tumor. Treatment may be
administered to a subject who does not exhibit signs of a disease,
disorder, and/or condition and/or to a subject who exhibits only
early signs of a disease, disorder, and/or condition for the
purpose of decreasing the risk of developing pathology associated
with the disease, disorder, and/or condition such as for example
Parkinson's Disease.
[0717] Unmodified: As used herein, "unmodified" refers to any
substance, compound or molecule prior to being changed in any way.
Unmodified may, but does not always, refer to the wild type or
native form of a biomolecule or entity. Molecules or entities may
undergo a series of modifications whereby each modified substance,
compound, molecule or entity may serve as the "unmodified" starting
molecule for a subsequent modification.
[0718] Vector: As used herein, a "vector" is any molecule or moiety
which transports, transduces or otherwise acts as a carrier of a
heterologous molecule. Vectors of the present disclosure may be
produced recombinantly and may be based on and/or may comprise
adeno-associated virus (AAV) parent or reference sequence. Such
parent or reference AAV sequences may serve as an original, second,
third or subsequent sequence for engineering vectors. In
non-limiting examples, such parent or reference AAV sequences may
comprise any one or more of the following sequences: a
polynucleotide sequence encoding a polypeptide or
multi-polypeptide, which sequence may be wild-type or modified from
wild-type and which sequence may encode full-length or partial
sequence of a protein, protein domain, or one or more subunits of a
protein; a polynucleotide comprising a modulatory or regulatory
nucleic acid which sequence may be wild-type or modified from
wild-type; and a transgene that may or may not be modified from
wild-type sequence. These AAV sequences may serve as either the
"donor" sequence of one or more codons (at the nucleic acid level)
or amino acids (at the polypeptide level) or "acceptor" sequences
of one or more codons (at the nucleic acid level) or amino acids
(at the polypeptide level).
[0719] Viral construct vector: As used herein, a "viral construct
vector" is a vector which comprises one or more polynucleotide
regions encoding or comprising Rep and or Cap protein.
[0720] Viral construct expression vector: As used herein, a "viral
construct expression vector" is a vector which comprises one or
more polynucleotide regions encoding or comprising Rep and or Cap
that further comprises one or more polynucleotide regions encoding
or comprising components for viral expression in a viral
replication cell.
[0721] Viral genome: As used herein, a "viral genome" is a
polynucleotide encoding at least one inverted terminal repeat
(ITR), at least one regulatory sequence, and at least one payload.
The viral genome is derived by replication of a payload construct
from the payload construct expression vector. A viral genome
encodes at least one copy of the payload construct.
EQUIVALENTS AND SCOPE
[0722] Those skilled in the art will recognize, or be able to
ascertain using no more than routine experimentation, many
equivalents to the specific embodiments in accordance with the
disclosure described herein. The scope of the present disclosure is
not intended to be limited to the above Description, but rather is
as set forth in the appended claims.
[0723] In the claims, articles such as "a," "an," and "the" may
mean one or more than one unless indicated to the contrary or
otherwise evident from the context. Claims or descriptions that
include "or" between one or more members of a group are considered
satisfied if one, more than one, or all of the group members are
present in, employed in, or otherwise relevant to a given product
or process unless indicated to the contrary or otherwise evident
from the context. The disclosure includes embodiments in which
exactly one member of the group is present in, employed in, or
otherwise relevant to a given product or process. The disclosure
includes embodiments in which more than one, or all of the group
members are present in, employed in, or otherwise relevant to a
given product or process.
[0724] It is also noted that the term "comprising" is intended to
be open and permits but does not require the inclusion of
additional elements or steps. When the term "comprising" is used
herein, the term "consisting of" is thus also encompassed and
disclosed.
[0725] Unless otherwise defined, all technical and scientific terms
used herein have the same meaning as commonly understood by one of
ordinary skill in the art to which this disclosure belongs. Methods
and materials are described herein for use in the present
disclosure; other, suitable methods and materials known in the art
can also be used.
[0726] Where ranges are given, endpoints are included. Furthermore,
it is to be understood that unless otherwise indicated or otherwise
evident from the context and understanding of one of ordinary skill
in the art, values that are expressed as ranges can assume any
specific value or subrange within the stated ranges in different
embodiments of the disclosure, to the tenth of the unit of the
lower limit of the range, unless the context clearly dictates
otherwise.
[0727] In addition, it is to be understood that any particular
embodiment of the present disclosure that falls within the prior
art may be explicitly excluded from any one or more of the claims.
Since such embodiments are deemed to be known to one of ordinary
skill in the art, they may be excluded even if the exclusion is not
set forth explicitly herein. Any particular embodiment of the
compositions of the disclosure (e.g., any nucleic acid or protein
encoded thereby; any method of production; any method of use; etc.)
can be excluded from any one or more claims, for any reason,
whether or not related to the existence of prior art.
[0728] All cited sources, for example, references, publications,
databases, database entries, and art cited herein, are incorporated
into this application by reference, even if not expressly stated in
the citation. In case of conflicting statements of a cited source
and the instant application, the statement in the instant
application shall control.
[0729] Section and table headings are not intended to be
limiting.
EXAMPLES
Example 1. Design of Payloads: AADC Polynucleotides
[0730] AADC polynucleotides are designed to comprise at a minimum a
nucleic acid sequence encoding an AADC protein.
[0731] Once designed, the sequence is engineered or synthesized or
inserted in a plasmid or vector and administered to a cell or
organism. Suitable plasmids or vectors are any which transduce or
transfect the target cell.
[0732] Adeno-associated viral (AAV) particles may be used.
[0733] Administration results in the processing of the AADC
polynucleotide to generate the AADC protein which alters the
etiology of the disease, in this case Parkinson's Disease. In one
non-limiting example, plasmids containing an AADC polynucleotide of
the disclosure have a CMV promoter and encode AADC. In some
embodiments the open reading frame of the AADC protein mRNA is
codon optimized.
[0734] AADC polynucleotides, listed ITR to ITR, suitable for use in
a AAV particles include those in Table 2.
TABLE-US-00003 TABLE 2 ITR to ITR AADC polynucleotides Construct
SEQ ID NO AADC Polynucleotide 979
[0735] The start and stop positions of various regions of the AADC
polynucleotides are given are relative to the ITR to ITR AADC
polynucleotides described in Table 2. In Table 3, ITR is inverted
terminal repeat, MCS is multiple cloning site, CMV is
cytomegalovirus, Ie1 is immediate-early 1, hBglobin is human
beta-globin, AADC is region encoding the AADC polypeptide, and
poly(A) is the polyadenylation signal.
TABLE-US-00004 TABLE 3 Component regions of AADC polynucleotides
AADC Polynucleotide (SEQ ID NO: 979) Length SEQ ID NO Region Start
Stop of Region of region 5' ITR 1 141 141 980 MCS 189 206 18 981
CMV enhancer 213 515 303 982 CMV promoter 516 719 204 983 Ie1 exon
1 734 867 134 984 Ie1 intron partial 868 899 32 985 hBglobin intron
2 900 1246 347 986 hBglobin exon 3 1247 1299 53 987 AADC 1338 2777
1440 988 MCS 2820 2837 18 989 Poly(A) 2838 3314 477 990 3' ITR 3386
3526 141 991
Example 2. Design of AADC Polynucleotides to Treat Parkinson's
Disease
[0736] AADC polynucleotides are designed to comprise at a minimum a
nucleic acid sequence encoding an AADC protein.
[0737] Once designed, the sequence is engineered or synthesized or
inserted in a plasmid or vector and administered to a cell or
organism. Suitable plasmids or vectors are any which transduce or
transfect the target cell.
[0738] Adeno-associated viral (AAV) particles may be used.
[0739] Administration results in the processing of the AADC
polynucleotide to generate the AADC protein which Alters the
Etiology of the Disease, in this Case Parkinson's Disease.
Example 3. Administration of AAV Particles
[0740] AAV particles are infused into the substantia nigra, and in
particular, the substantia nigra pars compacta (SNpc) and ventral
tegmental area (VTA) of patients having Parkinson's Disease and
identified as qualified for treatment according to methods known in
the art.
[0741] One method of administration contemplated for use in the
methods described herein is real-time convection-enhanced delivery
(RCD) of AAV particle compositions by co-infusion of gadoteridol (a
magnetic resonance (MR) contrast agent) and T1 or T2 magnetic
resonance imaging (MRI), which can predict areas of subsequent AADC
gene expression. As described in Richardson, et al., 2011, the
accuracy of cannula placement and initial infusate distribution may
be safely determined by saline infusion without significantly
altering the subsequent distribution of the tracer agent
(Richardson, et al., 2011, Neurosurgery, 69(1): 154-163). T2 RCD
provides detection of intraparenchymal convection-enhanced delivery
in the uninjured brain and may predict subsequent distribution of a
transgene after AAV particle infusion. Subjects undergo saline
infusion/T2 acquisition, immediately followed by gadoteridol
infusion/T1 acquisition in the putamen and brainstem. Distribution
volumes and spatial patterns are analyzed. Gadoteridol and
AAV-encoded AADC are co-infused under alternating T2/T1 acquisition
in the thalamus, and hyperintense areas are compared with areas of
subsequent transgene expression. Ratios of distribution volume to
infusion volume are expected to be similar between saline and
gadoteridol RCD. Spatial overlap should correlate well between T2
and T1 images. The second infusate will follow a spatiotemporal
pattern similar to that of the first, filling the target area
before developing extra-target distribution. Areas of AADC
expression should correlate well with areas of both T1 and T2
hyperintensity observed during RCD (Richardson, et al., 2011,
Neurosurgery, 69(1):154-163).
[0742] Convection-enhanced delivery (CED) of macromolecules
directly into the brain parenchyma has been known for over two
decades. CED is a term that denotes the use of a pressure gradient
to generate bulk flow within the brain parenchyma, i.e. convection
of macromolecules within the interstitial fluid driven by infusing
a solution through a cannula placed directly in the targeted
structure. This method allows therapeutic agents to be homogenously
distributed through large volumes of brain tissue by bypassing the
blood brain barrier and surpassing simple diffusion (Richardson, et
al., 2011, Stereotact. Funct. Neurosurg. 89:141-151).
[0743] Salegio, et al. recently demonstrated the distribution of
nanoparticles of different sizes, including micelles (.about.15 nm
in size), AAV (.about.20-25 nm) and liposomes (.about.65 nm),
within the CNS of rodents and NHPs (Salegio et al., 2014, Frontiers
in Neuroanatomy, vol. 8, article 9: pp. 1-8). Simple injections
cannot engage the perivascular system, and specialized infusion
cannulae are required, enabling constant pressures to be exerted at
the tip of the cannula such that the interstitial hydrostatic
pressure is exceeded and infusate can flow out into the tissue.
Simple needles generate significant reflux; thus, reflux-resistant
cannulas have been developed to counter this tendency. The advent
of platforms for MRI-guided convection-enhanced infusions further
refined understanding of the mechanics of perivascular flow, and it
was demonstrated that perivascular distribution of liposomes was
linear with respect to time, the slope of the curve was increased
in myelinated regions, and cessation of infusion prevented further
expansion in the volume of distribution. (Richardson, et al., 2011,
Stereotact. Funct. Neurosurg. 89:141-151; Salegio et al., 2014,
Frontiers in Neuroanatomy, vol. 8, article 9: pp. 1-8).
[0744] Intraparenchymal rAAV injections are known to result in
robust but relatively local transduction. Such local delivery
methods are advantageous when attempting gene therapy for
neurological disorders that result from neuropathology that is
localized to a specific anatomical region or anatomical circuitry
such as in the case of Parkinson's disease. However, in treatments
requiring more widespread CNS transduction, intraparenchymal
injections are impractical. Treatment of neurological disorders
attributable to inborn errors of metabolism and/or single-gene
defects, or those that affect motor neurons of the spinal cord can
require transduction of large proportions of the brain or spinal
cord, respectively. Development of less invasive trans-BBB delivery
methods for vectors is an extremely important endeavor. Numerous
attempts to use molecules that are known to interact with various
active transport mechanisms (probably receptor-mediated) to convey
proteins across the BBB have been reported with varying results.
Given the large number of AAV serotypes available, one or more
serotypes may bind a cell-entry receptor capable of transporting
the AAV capsid across the BBB (Manfredsson, et al., 2009, "AAV9: a
potential blood-brain barrier buster." Molecular Therapy
17(3):403-405).
Vector and Stereotaxic Infusion
[0745] A stereotactic approach may be used to surgically deliver
the AADC polynucleotides. Although individuals with AADC deficiency
lack epinephrine and norepinephrine, these patients should maintain
stable blood pressure and heart rates during the surgery. There
should be no notable intracerebral hemorrhages in the postoperative
computed tomography (CT) or MRI scans. The needle tracts, as shown
on the MRI scans, should show accurate injection into the
substantia nigra pars compacta (SNpc) and ventral tegmental area
(VTA). The patients will be discharged from the hospital about one
week after the surgery (Hwu, W. L., et al., 2012. Gene therapy for
aromatic L-amino acid decarboxylase deficiency. Sci. Transl. Med.
Vol. 4, 134ra61).
[0746] Subjects of treatment receive the AAV-vector composition
vector, safely delivered to substantia nigra pars compacta (SNpc)
and ventral tegmental area (VTA) via bilateral infusions, or
alternatively, intrastriatally (into the caudate nucleus and
putamen), or into the subthalamic nucleus (STN), for example
optionally using the FDA-approved SMARTFLOW.RTM. neuroventricular
cannula (SurgiVision, Inc.) specifically designed for clinical
application, with or without the aid of the CLEARPOINT.RTM. system
to help the treating neurosurgeon(s) target and observe the
delivery of the therapeutic agent in the brain (See, for example,
San Sebastian, et al., 2014, Mol. Ther. Methods Clin. Dev. 3:
14049; See, for example, Feng and Maguire-Zeiss, 2010, CNS Drugs
24(3):177-192).
[0747] For example, during the surgery, two target points are
determined in the substantia nigra pars compacta (SNpc) and ventral
tegmental area (VTA) that are sufficiently separated from each
other in dorsolateral directions and identified on a magnetic
resonance image. One burr hole is trepanned in each side of the
cranial bone, through which the vector is injected into the two
target points via the two-track insertion route. The
AAV-vector-containing solution is prepared to a concentration of
1.5.times.10.sup.12 vector genome/ml, and 50 .mu.l per point of the
solution is injected at 1 .mu.l/min; each patient receives
3.times.10.sup.11 vector genome of the AAV-vector construct.
[0748] Neutralizing antibody titers against AAV2 are determined by
measuring .beta.-galactosidase activities in HEK293 cells
transduced with 5.times.10.sup.3 vector genome/cell of AAV2 vectors
expressing (3-galactosidase in various dilutions of sera.
PET
[0749] The AADC expression level in the substantia nigra are
assessed on PET imaging with FMT six days before surgery and at
one- and six-months after gene transfer. All patients cease taking
dopaminergic medications 18 hours before PET and take 2.5 mg/kg of
carbidopa orally one hour before FMT injection. Subsequently, 0.12
mCi/kg of FMT in saline is infused into an antecubital vein, and a
90-minute dynamic acquisition sequence is obtained. The PET and
magnetic resonance imaging data are co-registered with a fusion
processing program (Syntegra; Philips, Amsterdam, The Netherlands)
to produce the fusion images. Radioactivities within volumes of
interest drawn in the nigrostriatal pathway are calculated between
80 and 90 minutes after tracer injection. A change in nigrostriatal
pathway FMT uptake from baseline to 24 weeks is assessed using the
substantia nigra to striatal ratio of radioactivity.
Statistical Analysis
[0750] Values at baseline and 6 months after gene transfer are
compared using Student's t-test (paired analyses). A two-sided P
value <0.05 is taken to indicate significant differences.
Two-way analysis of variance with Bonferroni correction of P values
is used for the short-duration response to levodopa. (See, for
example, Muramatsu, et al., 2010, "A phase I study of aromatic
L-amino acid decarboxylase gene therapy for Parkinson's disease."
Mol. Ther. 18:1731-1735).
[0751] Safety and tolerability of bilateral administration of
AAV-vector compositions using real-time image-guided infusion into
the brains of Parkinson's Disease subjects may be monitored for up
to or after 9 months post-surgery. Broad coverage of targeted areas
(substantia nigra pars compacta (SNpc) and ventral tegmental area
(VTA)) and widespread AADC protein distribution in the striatum
should be achieved without inducing any adverse effects.
[0752] Changes in Growth and Motor Skills:
[0753] The patients should gain weight and exhibit improvement in
their motor scores after gene transfer, within a year,
post-treatment. Weight will be measured at 3 to 6 months after gene
transfer. All patients initially should have raw scores of zero on
the Alberta Infant Motor Scale (AIMS) and very low raw scores for
the Peabody Developmental Motor Scale, Second Edition (PDMS-II).
After the gene transfer, all of the patients should show continuous
increases in their raw scores on these two scales, which indicates
that their motor functions have improved. The Comprehensive
Developmental Inventory for Infants and Toddlers (CDIIT) covers
both cognition and motor development. All of the patients should
show low raw CDIIT scores before gene transfer, and the subsequent
increase in scores demonstrate improvement in both motor and
cognitive functions.
Subjective Improvements after Gene Transfer To document the
symptoms that are more difficult to quantify, spouses, guardians or
caretakers of the patients are asked to fill out a questionnaire at
the end of the study. The symptoms of the oculogyric crises should
lessen, and eye deviations and sleep disruptions, for example, are
some mild symptoms of the oculogyric crises that may remain after
gene therapy. Subjects may experience increased emotional
stability, and/or some improvements in sweating and hyperthermia (a
common manifestation of body temperature instability in hot
weather). There should be no detectable abnormality in heart rate
variability as assessed by 24-hour Holter monitoring either before
or after gene transfer. Before gene therapy, patients that were
bedridden and showed little spontaneous movement may exhibit less
severe ptosis (drooping of the upper eyelid) one to two weeks after
the gene transfer. According to previous studies, dyskinesia may
occur one month after gene transfer, but upon observation of a
decrease in dyskinesia, motor development should start (Hwu, W. L.,
et al., 2012. Gene therapy for aromatic L-amino acid decarboxylase
deficiency. Sci. Transl. Med. Vol. 4, 134ra61). Subjects may
exhibit increased head control after three months, sitting with
support after six to nine months, sitting up from the prone
position after thirteen months, and holding toys and standing with
support sixteen months after the gene transfer, for example.
Anti-AAV2 antibodies should be negative in the patients before gene
therapy, and the titers may increase slightly after gene
transfer.
PET Scans and CSF Analyses
[0754] PET scans and CSF analyses are completed for the treated
patients. Six months after gene transfer, PET scans should reveal
that uptake of 6-[18F] fluorodopa (FDOPA) increase from baseline in
the combined (right and left) treatment sites. The CSF analysis
should reveal increases in the levels of homovanillic acid (HVA, a
metabolite of dopamine) and 5-hydroxyindoleacetic acid (HIAA, a
metabolite of serotonin). However, the levels of L-DOPA and
3-O-methyldopa may remain elevated (Hwu, W. L., et al., 2012. Gene
therapy for aromatic L-amino acid decarboxylase deficiency. Sci.
Transl. Med. Vol. 4, 134ra61).
Example 4. Administration of AADC Polynucleotides
[0755] AAV particle compositions are infused into the putamen of
patients having Parkinson's Disease using the administration
methods described in Example 3. The dose, number of patients and
volume are outlined in Table 4.
TABLE-US-00005 TABLE 4 Study Design Number of Study No. Patients
Dose Volume 1 6 .sup. 3 .times. 10.sup.11 vg 100 ul per putamen 2 6
.sup. 9 .times. 10.sup.11 vg 300 ul per putamen 3 10 2.3 .times.
10.sup.11 vg 100 ul per putamen 4 10 7.5 .times. 10.sup.11 vg 100
ul per putamen 5 5 7.5 .times. 10.sup.11 vg 450 ul per putamen 6 Up
to 20 1.4 .times. 10.sup.12 vg Up to 900 ul per putamen 7 Up to 20
4.8 .times. 10.sup.12 vg Up to 900 ul per putamen 8 Up to 20 8.8
.times. 10.sup.12 vg Up to 900 ul per putamen
[0756] During the course of the study the safety and tolerability
of the infusion of the AADC polynucleotide-containing recombinant
adeno-associated virus (AAV) vector compositions in human patients
diagnosed with Parkinson's Disease is evaluated. Patients are
evaluated preoperatively and monthly postoperatively for six
months, using multiple measures, including the Global Systonia
Scale (GDS) (see Comella, et al., 2003, Movement Disorders,
18(3):303-312), L-DOPA challenge test, UPDRS scores, motor state
diaries, and laboratory tests. Using diaries that separate the day
into half-hour segments, the caregivers of the patients will record
their mobility during the four days before admission and for
another four days at six months after admission to the study site.
The patient caregivers are trained to rate subject's condition as
sleeping, immobile, mobile without troublesome dyskinesias, or
mobile with troublesome dyskinesias. The total number of hours
spent in each of these categories is calculated, and the
differences between the baseline and the six-month scores are
compared between the groups. The short-duration response to
levodopa is evaluated at baseline and 6 months after gene transfer;
subjects take 100 mg of levodopa orally with 25 mg benserazide
after 20 hours without dopaminergic medication. Motor symptoms
based on GDS and plasma levodopa concentrations are assessed at
baseline and 30 minutes, 1, 2, 3, and 4 hours after levodopa intake
(See, for example, Muramatsu, et al., 2010, "A phase I study of
aromatic L-amino acid decarboxylase gene therapy for Parkinson's
disease." Mol. Ther. 18:1731-1735).
[0757] While the present disclosure has been described at some
length and with some particularity with respect to the several
described embodiments, it is not intended that it should be limited
to any such particulars or embodiments or any particular
embodiment, but it is to be construed with references to the
appended claims so as to provide the broadest possible
interpretation of such claims in view of the prior art and,
therefore, to effectively encompass the intended scope of the
disclosure.
[0758] All publications, patent applications, patents, and other
references mentioned herein are incorporated by reference in their
entirety. In case of conflict, the present specification, including
definitions, will control. In addition, section headings, the
materials, methods, and examples are illustrative only and not
intended to be limiting.
Sequence CWU 0 SQTB SEQUENCE LISTING The patent application
contains a lengthy "Sequence Listing" section. A copy of the
"Sequence Listing" is available in electronic form from the USPTO
web site
(http://seqdata.uspto.gov/?pageRequest=docDetail&DocID=US20190060425A1).
An electronic copy of the "Sequence Listing" will also be available
from the USPTO upon request and payment of the fee set forth in 37
CFR 1.19(b)(3).
0 SQTB SEQUENCE LISTING The patent application contains a lengthy
"Sequence Listing" section. A copy of the "Sequence Listing" is
available in electronic form from the USPTO web site
(http://seqdata.uspto.gov/?pageRequest=docDetail&DocID=US20190060425A1).
An electronic copy of the "Sequence Listing" will also be available
from the USPTO upon request and payment of the fee set forth in 37
CFR 1.19(b)(3).
* * * * *
References