This application from VIROVEK INCORPORATION seeks to patent a method of healing cancer! 10 minutes of your time can help narrow this US patent application before it becomes a patent.
QUESTION - Have you seen anything that was published before 2013/08/15 that discusses:
- The use of a insect cell for a baculovirus, a retrovirus, an adenovirus, an adeno-associated virus production.
- The use of a toxic gene for mammals not expressed in insect cells.
If you've ever seen anything like this before, please submit evidence of prior art as an answer to this question.
EXTRA CREDIT - Include a reference to anything that meets all of the criteria to the question above AND ALSO the use of a gene in a viral vector expressed in a insect cell or method of obtaining it.
TITLE: VECTORS HARBORING TOXIC GENES, METHODS AND USES THEREFOR
Summary: The present disclosure describes nucleic acids, and viruses comprising such nucleic acids, for growing a toxic gene in an insect cell. These nucleic acids comprise a sequence encoding a toxic polypeptide, and an intron that interrupts the sequence, whereby the intron is spliced in mammalian cells but not in insect cells. Infection of mammalian cells but not insect cells with the nucleic acids or viruses can lead to expression of toxic levels of the toxic polypeptide in mammalian cells but not in insect cells. Viruses, such as an AAV or a baculovirus comprising a nucleic acid can be grown in insect cell lines in vitro and can be administered to a subject in need of therapy, such as a subject in need of cancer therapy.
Publication Number: US2014296324
Assignee: CHEN HAIFENG; VIROVEK INC
Prior Art Date: Seeking prior Art predating 16 Oct. 2012
Open for Challenge at USPTO: Open through 2 Apr. 2015
The claims verbatim:
- A nucleic acid comprising: a sequence encoding a toxic polypeptide; and an intron that interrupts the sequence, whereby the intron is
spliced in mammalian cells but not in insect cells to form an mRNA
that is translated to form cell-toxic levels of the toxic polypeptide
in mammalian cells but not in insect cells.
- A nucleic acid in accordance with claim 1, wherein the toxic polypeptide is selected from the group consisting of diphtheria toxin
(DT-A), barnase, ricin, abrin, and Pseudomonas exotoxin.
- A nucleic acid in accordance with claim 1, wherein the toxic polypeptide is diphtheria toxin (DT-A).
- A nucleic acid in accordance with claim 1, wherein the toxic polypeptide is a barnase polypeptide.
- A nucleic acid in accordance with claim 1, further comprising at least one expression element operably linked to the sequence.
- A nucleic acid in accordance with claim 5, wherein the expression element is selected from the group consisting of a promoter, an IRES,
an enhancer and a combination thereof.
- A nucleic acid in accordance with claim 5, wherein the expression element is selected from the group consisting of a CMV promoter, an
AFP promoter, an AFP enhancer, an hSURV promoter, a CXCR4 promoter, an
hTERT promoter, a COX2 promoter, and a CCKAR promoter.
- A nucleic acid in accordance with claim 5, wherein the expression element is selected from the group consisting of an hSURV promoter and
a CXCR4 promoter.
- A nucleic acid in accordance with any one of claims 1-8, wherein the intron is an artificial intron.
- A nucleic acid in accordance with any one of claims 1-8, wherein, the intron is a human growth hormone intron.
- A nucleic acid in accordance with any one of claims 1-8, wherein the intron is a SV40 large T-antigen intron.
- A nucleic acid in accordance with any one of claims 1-8, comprising, in 5′ to 3′ order, a first ITR, a promoter, a first
portion of a toxic gene, an intron, a second portion of the toxic
gene, a polyadenylation signal, and a second ITR.
- A nucleic acid in accordance with claim 13, wherein the first ITR is an AAV ITR.
- A nucleic acid in accordance with claim 13, wherein the second ITR is an AAV ITR.
- A nucleic acid in accordance with claim 13, wherein the promoter is a CMV promoter.
- A nucleic acid in accordance with claim 13, wherein the promoter is a hSURV promoter.
- A nucleic acid in accordance with claim 13, wherein the promoter is a hCXCR4 promoter.
- A nucleic acid in accordance with claim 13, wherein the promoter is a hAFP promoter.
- A nucleic acid in accordance with claim 13, wherein the toxin gene encodes DT-A.
- A nucleic acid in accordance with claim 13, wherein the toxic gene encodes Barnase.
- A viral vector comprising the nucleic acid of any one of claims 1-8.
- A viral vector in accordance with claim 22, wherein the vector is selected from the group consisting of a baculovirus, a retrovirus, an
adenovirus, an adeno-associated virus (AAV) and a combination thereof.
- A viral vector in accordance with claim 22, wherein the vector is a baculovirus, an adeno-associated virus (AAV), or a combination
thereof.
- A viral vector in accordance with claim 22, wherein the vector is an adeno-associated virus (AAV).
- A viral vector in accordance with claim 22, wherein the vector is an AAV2.
- A viral vector in accordance with claim 22, wherein the vector is an AAV9.
- A viral vector in accordance with claim 22, wherein the vector comprises an ITR.
- An insect cell in vitro comprising the nucleic acid of any one of claims 1-8.
- An insect cell in accordance with claim 29, wherein the cell is selected from the group consisting of a Trichoplusia ni BTI-Tn-5B1-4
cell, a Spodoptera frugiperda Sf9 cell and a Spodoptera frugiperda
Sf21 cell.
- An insect cell in accordance with claim 29, wherein the nucleic acid is comprised by a viral genome.
- An insect cell in accordance with claim 29, wherein the nucleic acid is comprised by a virus.
- An insect cell in accordance with claim 29, wherein the cell is stably transformed with the nucleic acid of any one of claims 1-8.
- A cell culture comprising: a plurality of cells of claim 29; and a culture medium.
- A cell culture in accordance with claim 34, wherein unconcentrated culture medium comprises greater than 109 viral genomes/ml.
- A cell culture in accordance with claim 34, wherein unconcentrated culture medium comprises greater than 1010 viral genomes/ml.
- A cell culture in accordance with claim 34, wherein unconcentrated culture medium comprises greater than 1011 viral genomes/ml.
- A cell culture in accordance with claim 34, wherein unconcentrated culture medium comprises greater than 1012 viral genomes/ml.
- A cell culture in accordance with claim 34, wherein unconcentrated culture medium comprises a baculovirus greater than 106 PFU/ml.
- A cell culture in accordance with claim 34, wherein unconcentrated culture medium comprises a baculovirus greater than 107 PFU/ml.
- A cell culture in accordance with claim 34, wherein unconcentrated culture medium comprises a baculovirus greater than 108 PFU/ml.
- A cell culture in accordance with claim 34, wherein unconcentrated culture medium comprises a baculovirus greater than 109 PFU/ml.
- A method of growing a vector comprising a toxic gene in vitro, comprising: providing a cell culture comprising insect cells;
infecting or transfecting the cells with a nucleic acid of any one of
claims 1-8; and incubating the cells under conditions suitable for
virus production.
- A method of treating a cancer, comprising administering to a subject in need of therapy a therapeutically effective amount of the
nucleic acid of any one of claims 1-8.
- A method of treating a cancer, comprising administering a therapeutically effective amount of an adeno-associated virus
comprising the nucleic acid of any one of claims 1-8 to a subject in
need of therapy.
- A method of treating a cancer in accordance with claim 45 wherein the cancer is selected from the group consisting of prostate cancer,
breast cancer, brain tumor, cervical cancer and nasopharyngeal
carcinoma.
- Use of a nucleic acid of any one of claims 1-8 for the treatment of a cancer.
- A nucleic acid in accordance with any one of claims 1-8 for use in the treatment of a cancer.
- Use of a nucleic acid of any one of claims 1-8 in the manufacture of a medicament for treatment of a cancer.
- Use of a viral vector of claim 22 for the treatment of a cancer.
- The viral vector of claim 22 for use in the treatment of a cancer.
- Use of a viral vector of claim 22 in the manufacture of a medicament for treatment of a cancer.
- A method of producing AAV comprising a toxic gene in vitro, comprising: providing a cell culture comprising insect cells;
infecting the cells with a virus vector of claim 24 and with
Bac-inCap-inRep for at least about 1 day; and lysing the cells to form
a lysate comprising the AAV.
- A method in accordance with claim 53, wherein the infecting the cells for at least about 1 day comprises infecting the cells for at
least 2 days.
- A method in accordance with claim 53, wherein the infecting the cells for at least about 1 day comprises infecting the cells for at
least 3 days.
- A method in accordance with claim 53, wherein the infecting the cells for at least about 1 day comprises infecting the cells for about
3 days.
- A method in accordance with claim 53, wherein the infecting the cells for at least about 1 day comprises infecting the cells for at
least 4 days.
- A method in accordance with claim 53, wherein the infecting the cells for at least about 1 day comprises infecting the cells for at
least 5 days.
- A method in accordance with claim 53, wherein the infecting the cells for at least about 1 day comprises infecting the cells for up to
about 5 days.
- A method in accordance with claim 53, wherein the infecting the cells for at least about 1 day comprises infecting the cells for up to
about 6 days.
- A method in accordance with claim 53, wherein the infecting the cells for at least about 1 day comprises infecting the cells for up to
about 7 days.
- A method in accordance with claim 53, further comprising digesting the cellular debris with benzonase.
- A method in accordance with claim 53, wherein the AAV is AAV2.
- A method in accordance with claim 53, wherein the AAV is AAV9.
- A method in accordance with claim 53, wherein the insect cells are selected from the group consisting of Trichoplusia ni BTI-Tn-5B1-4
cells, Spodoptera frugiperda Sf9 cells, Spodoptera frugiperda Sf21
cells, and any combination thereof.
- A method in accordance with claim 53, wherein the insect cells are Spodoptera frugiperda Sf9 cells.
Here there is an image how the claims are related (from the espacenet web):
I think it might infringe many other patents. My concern is about using some of the information from these patent in other diseases. Also what does here the toxic gene mean?
Note: It is my first post here, I might be confused but I followed this guide and it might be not really what I should do. If I can improve my question, please let me know.