+

WO1996039416A1 - Therapie genique de l'adenome hypophysaire et d'autres troubles endocriniens - Google Patents

Therapie genique de l'adenome hypophysaire et d'autres troubles endocriniens Download PDF

Info

Publication number
WO1996039416A1
WO1996039416A1 PCT/US1996/008191 US9608191W WO9639416A1 WO 1996039416 A1 WO1996039416 A1 WO 1996039416A1 US 9608191 W US9608191 W US 9608191W WO 9639416 A1 WO9639416 A1 WO 9639416A1
Authority
WO
WIPO (PCT)
Prior art keywords
pituitary
cells
gene
dopamine
tumor
Prior art date
Application number
PCT/US1996/008191
Other languages
English (en)
Inventor
Andrew Freese
Eugene S. Flamm
Peter J. Snyder
Thomas Gennarelli
Matthew J. During
Original Assignee
The Trustees Of The University Of Pennsylvania
Yale University
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by The Trustees Of The University Of Pennsylvania, Yale University filed Critical The Trustees Of The University Of Pennsylvania
Priority to AU60282/96A priority Critical patent/AU6028296A/en
Publication of WO1996039416A1 publication Critical patent/WO1996039416A1/fr

Links

Classifications

    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N9/00—Enzymes; Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating or purifying enzymes
    • C12N9/0004—Oxidoreductases (1.)
    • C12N9/0071—Oxidoreductases (1.) acting on paired donors with incorporation of molecular oxygen (1.14)
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K48/00—Medicinal preparations containing genetic material which is inserted into cells of the living body to treat genetic diseases; Gene therapy

Definitions

  • This invention relates to the field of mammalian gene transfer, tumors of the hypothalamus-pituitary axis, and endocrine disorders. More particularly, methods are provided for delivering of genes that directly or indirectly modulate pituitary tumor and gland physiology.
  • prolactinomas lactotroph adenomas
  • the mainstay of therapy for prolactinomas currently focuses on the use of the ergot derivative, bromocriptine.
  • the primary physiological prolactin inhibitory factor is dopamine.
  • Bromocriptine a dopamine receptor agonist, apparently acts by suppressing the secretion of prolactin and also retarding the division of prolactin-secreting cells and growth of prolactinomas.
  • bromocriptine therapy has several problems. Some tumors are resistant to this therapy and continue to grow and secrete abnormal amounts of prolactin. I. Pellegrini et al., Horm. Res. 1989, 31, 19-23. In addition, relatively high doses must be continued on a daily, multiple dose basis as cessation of treatment results in the tumor resuming its previous abnormal secretory and growth pattern. Many patients also find the side effects of bromocriptine unacceptable and cease treatment. New dopaminergic drugs with longer lasting action than bromocriptine, have been reported. However, these new drugs still require continued treatment with attendant side-effects. M.S. Venetikou et al., Acta Endocr ⁇ nol (Copenhagen) 1987, 116, 287-92.
  • GHRP growth hormone releasing protein
  • prolactinomas in men are frequently clinically silent until they reach a macroadenoma size, prolactinomas are more difficult to treat in male patients. Furthermore, once the tumors reach this large size, they can invade relatively inaccessible areas, including the cavernous sinus, rendering surgical removal oftentimes impossible.
  • a method for transducing gene sequences encoding other hormones, receptors or modulators into the pituitary gland can be used to alter hormone secretion by and growth patterns of other pituitary adenomas, as well as modulate pituitary function in other disease conditions.
  • An object of the present invention is to provide a method of delivering at least one gene encoding a dopamine synthetic enzyme to pituitary cells which comprises contacting pituitary cells with a viral vector comprising this gene.
  • Another object of the present invention is to provide a method of modulating pituitary tumor physiology which comprises contacting pituitary tumor cells with a viral vector comprising at least one gene capable of encoding a dopamine synthetic enzyme which modulates pituitary tumor physiology.
  • Another object of the present invention is to provide viral vectors containing at least one gene encoding a dopamine synthetic enzyme which is capable of modulating pituitary tumor physiology, including, but not limited to, genes encoding tyrosine hydroxylase, amino acid decarboxylase (or dopa decarboxylase) and GTP cyclohydroxylase (responsible for production of the tyrosine hydroxylase cofactor, tetrahydrobiopterin).
  • Another object of the present invention is to provide viral vectors containing at least one gene encoding other factors that modulate pituitary gland or tumor physiology, including, but not limited to, somatostatin, growth hormone releasing factor (GHRF), activin, follistatin, (luteinizing hormone/follicle stimulating hormone (LH/FSH), corticotropin releasing factor (CRF), corticotropin releasing inhibitory factor (CRIF), adrenocorticotrophic hormone (ACTH), growth hormone (GH), gonadotroph releasing hormone (GNRH), thyroid stimulating hormone (TSH) and other proteins or enzymes endogenous to the hypothalamus-pituitary axis.
  • somatostatin growth hormone releasing factor (GHRF), activin, follistatin, (luteinizing hormone/follicle stimulating hormone (LH/FSH), corticotropin releasing factor (CRF), corticotropin releasing inhibitory factor (CRIF), adrenocorticotrophic hormone (ACTH),
  • Another object of the present invention is to provide viral vectors containing at least one gene encoding receptors for factors that modulate pituitary gland or tumor physiology, including, but not limited to, dopamine receptors (particularly the D2 receptor), somatostatin receptors, activin and follistatin receptors, as well as other receptors for proteins and compounds enumerated above.
  • dopamine receptors particularly the D2 receptor
  • somatostatin receptors particularly the D2 receptor
  • activin and follistatin receptors as well as other receptors for proteins and compounds enumerated above.
  • Another object of the present invention is to provide viral vectors containing two or more separate genes that each encode a separate protein that may alter pituitary gland or tumor physiology, including, but not limited to, the combination of tyrosine hydroxylase and amino acid decarboxylase, or tyrosine hydroxylase and somatostatin.
  • Figure 1 provides schematics of the viral vectors used in the gene transduction experiments.
  • Figure 1A shows a modified human herpes simplex virus-1 vectors, pHSVth, containing a human tyrosine hydroxylase II gene under the control of the herpes immediate early 4/5 promoter (HSV EI-4/5).
  • Figure IB shows an adenovirus vector containing a human tyrosine hydroxylase gene under control of an RSV promoter.
  • Figure 1C shows an adeno-associated virus vector containing a tyrosine hydroxylase gene.
  • Figure ID shows a bicistronic adeno-associated viral vector containing both the tyrosine hydroxylase gene and an amino-acid decarboxylase gene (the gene encoding the next enzyme in the dopamine biosynthetic pathway) in tandem.
  • Figure 2 is a schematic outline of the biosynthetic pathway of dopamine biosynthesis with the key intermediates and their corresponding enzymes indicated.
  • Figure 3 is the X-gal-based detection (blue cells) of ⁇ - galactosidase activity in human pituitary cells following transfection with either an AAV vector with the Lac Z gene as shown in Figure 3A or an Adenovirus vector with the Lac Z gene as shown in Figure 3B, demonstrating the first successful transfer of a transgene (Lac Z) encoding a foreign protein ( ⁇ -galactosidase) in human pituitary cells.
  • transgene Lac Z
  • FIG 3C negative controls (cells not transfected with the viral vector) do not show blue staining in the cells.
  • Figure 4 is the immunohistochemical detection of TH- transgene expression in the transduced human primary pituitary prolactinoma cells.
  • Adenovirus vector with the TH gene was used in Figure 4A
  • AAV vector with the TH and AADC genes and the Flag epitope was used in Figure 4B
  • control is shown in Figure 4C (absent staining).
  • Controls are mock transductions or transductions with vectors lacking the dopamine synthesis genes of pituitary and prolactinoma cells.
  • Figure 5 shows the inhibition of prolactin secretion from cells transduced with the adeno-associated virus containing both the TH and AADC genes.
  • the levels of prolactin are assayed in the cell culture media.
  • the percent of prolactin secretion by the adenoma cultures was statistically the same in both lower (depicted as lightly shaded bar) and higher (depicted as darker shaded bar) viral titer-treated wells (2xl0 5 and 5xl0 5 particles/ml).
  • the prolactin secretion (as a percentage of control) was the same in the viral vector-treated cultures as in those cultures exposed to supraphysiological doses of dopamine (depicted as filled bar), confirming that the vectors resulted in the same reduction of prolactin secretion as did supraphysiological doses of dopamine.
  • dopamine receptor analogues such as bromocriptine.
  • Figure 6 shows reduction in prolactin secretion by human primary prolactinoma cells in cultures transduced with the adenoviral vector containing the TH transgene, compared to control cells transduced with the non-physiological Lac Z gene (depicted as filled triangle).
  • the higher titer of vector (10 7 particles/ml, depicted as filled circle) resulting in a larger reduction than the lower titer of vector (10 5 particles/ml, depicted as square) .
  • Figure 7A shows dopamine secretion induced by an adenovirus viral vector containing the TH gene in human primary prolactinoma cells in culture compared to controls (the prolactinoma cells transduced with the Lac Z gene) .
  • Figure 7B shows L-dopa secretion induced by the same vector. In each situation, a marked increase of both L-dopa and dopamine secretion was noted.
  • Figure 8 provides cytotoxicity experiments in pituitary cells contacted with these viral vectors.
  • Viral vectors of the present invention are not cytotoxic to human pituitary cells in culture. Pituitary cells were counted in dishes several days following transfeetion with the vectors or control. Control constitutes the buffer, phosphate buffered saline (PBS). Vectors used were Adenovirus with the Lac Z gene and Adenovirus with the TH gene.
  • PBS phosphate buffered saline
  • viral vectors containing at least one gene encoding an enzyme involved in dopamine biosynthesis are useful in transducing the gene so that these enzymes are expressed in primary human prolactinoma tumor cells in culture.
  • the resultant in situ increase in dopamine has been shown to inhibit prolactin secretion from these prolactinoma cells.
  • the primary prolactin inhibitory factor is dopamine, it is believed that the introduction into the pituitary gland or hypothalamus of genes encoding enzymes involved in dopamine synthesis, which enhances dopamine synthesis and release and inhibits prolactin secretion, will modulate the physiology of prolactinomas.
  • enzymes are very important in the biosynthesis of dopamine.
  • these enzymes are referred to as "dopamine synthetic enzymes".
  • these enzymes include, but are not limited to, human tyrosine hydroxylase which catalyses a critical hydroxylation from the amino acid precursor tyrosine, and amino acid decarboxylase which catalyses a critical decarboxylation from the amino acid precursor tyrosine.
  • enzymes which the synthesis of cofactors needed in the synthesis of dopamine For example, tetrahydrobiopterin, is the cofactor for the rate limiting step in dopamine biosynthesis which is mediated by tyrosine hydroxylase.
  • viral vectors useful in the present invention can be routinely selected by those of skill in the art upon this disclosure. The vector selected should allow sufficient expression of the gene, while producing minimal viral gene expression. There should be minimal viral DNA replication and ideally no virus replication. In addition, recombination to produce new viral sequences and complementation to allow growth of the defective virus in an animal should be kept to a minimum.
  • Preferred viral vectors which can be used in the present invention include, but are not limited to, adenoviruses, adeno-associated viruses and herpesviruses.
  • Adenovirus-based vectors are well-suited for * gene therapy as they appear to be relatively safe and can be manipulated to encode a desired gene product, while at the same time, be inactivated in terms of their ability to replicate in a normally lytic viral life cycle.
  • Adenoviruses are able to infect quiescent cells. Expression of an adenovirus is achieved without integration of the viral DNA into the host cell chromosome thus alleviating concerns about insertional mutagenesis.
  • adenoviruses have been used as live enteric vaccines for many years with an excellent safety profile. Schwartz et al., Am. Rev. Respir. Di ⁇ . 1974, 109, 233-238.
  • Pseudo-adenoviruses may also be useful as vectors in the present invention.
  • PAV contains no potentially harmful viral genes, has a theoretical capacity for foreign material of nearly 36 kb, may be produced in reasonably high titers, and maintains the tropism of the parent adenovirus for dividing and non-dividing human target cell types.
  • PAVs comprise adenovirus inverted terminal repeats and the minimal sequences of a wild type adenovirus type 2 genome necessary for efficient replication and packaging by a helper virus and genetic material of interest.
  • Adeno-associated virus is a DNA-containing parvovirus that has not been directly linked to any human disease, even though the majority of Americans demonstrate seropositivity to AAV, AAV vectors can be generated which show minimal to no cytotoxicity and minimal or no protein synthesis other than the desired transgene. Samulski et al., J. Virol . 1989, 63, 3822-3828.
  • Herpes Simplex Virus Type-1 (HSV-1) vectors can be generated through recombinant techniques so that they contain transgenes under the control of selected viral or cellular promoters. By use of vectors with helper viruses or defective HSV-1 mutants, vectors can be produced that show safety and efficacy, with minimal or no cytotoxicity, viral replication and/or recombination. Freese et al., Biochem. Pharmacol . 1990, 40, 2189-2199.
  • Viral vectors containing at least one gene encoding other factors that modulate pituitary gland or tumor physiology including dopamine biosynthetic enzymes, dopamine receptors, somatostatin, somatostatin receptors, GHRF, activin, follistatin, activin or follistatin receptors, LH/FSH, CRF,
  • CRIF CRIF, ACTH, GH, GNRH, TSH and other proteins, enzymes or receptors endogenous to the hypothalamus-pituitary axis may also be used in accordance with the teachings of the invention.
  • the viral vector of the present invention comprises more than one gene encoding for different enzymes involved in dopamine biosynthesis.
  • the vector construct may contain genes encoding tyrosine hydroxylase and amino-acid decarboxylase. This construct represents an improvement over vectors encoding these two genes separately, as this tandem construct ensures that any transduced cell will receive both required enzymes simultaneously and not be subject to random binomial distribution of each gene delivered separately. Furthermore, this tandem construct is more likely to enhance the production of dopamine, rather than L-dopa.
  • Viral vectors of the present invention containing at least one gene encoding a dopamine synthetic enzyme are useful in delivering a gene encoding a dopamine synthetic enzyme to pituitary cells comprising contacting pituitary cells with a viral vector a selected gene encoding a dopamine synthetic enzyme.
  • These viral vectors can also be used in the modulation of pituitary tumor physiology, preferably prolactinomas.
  • Pituitary tumor cells are contacted with a viral vector containing a selected gene encoding a dopamine synthetic enzyme capable of modulating pituitary tumor physiology.
  • the viral vector comprises a gene encoding human tyrosine hydroxylase and a gene encoding amino acid decarboxylase.
  • the viral vectors of the present invention can also be used to modulate secretion of prolactin by pituitary cells in conditions such as hyperprolactinemia.
  • Hyperprolactinemia is a condition involving increased levels of prolactin in the blood. In women this condition is associated with amenorrhea and galactorrhea. In men, hyperprolactinemia is associated with hypogonadism and impotence. Individuals with hyperprolactinemia many times experience infertility. If associated with enlargement of the pituitary gland resulting from lactotroph hypertrophy, hyperplasia or adenoma formation, these individuals may also suffer from persistent headaches and vision disturbances. Viral vectors of the present invention can be used to decrease the levels of prolactin secreted from the pituitary cells.
  • the viral vectors of the present invention are administered to a mammal, preferably a human.
  • the vectors can be administered orally or parenterally including intravenously, intramuscularly, intraperitoneally, intranasally, subcutaneously and surgically.
  • a pharmaceutical vehicle suitable for injection such as a sterile aqueous solution or dispersion.
  • the mammal is monitored to detect improvement in outward manifestations. Prolactin levels are measured to determine when they approach or reach normal levels. Magnetic resonance imaging can be used to monitor any change in size of pituitary adenomas. Dose and duration of treatment is determined individually depending upon the degree and rate of improvement.
  • the viral vectors of the present invention provide the first demonstration of gene transduction of human cells of the hypothalamic-pituitary axis and use of gene transduction to modulate hypothalamic-pituitary axis physiology and pituitary tumor physiology, i.e., endocrine secretion.
  • These vectors provide a novel approach to gene therapy and, more specifically, tumor therapy; rather than causing tumor cell death or dysfunction, transducing genetic sequences as described in the present invention results in the production of proteins that either directly or indirectly modulate tumor physiology, hormone secretion and tumor size regression.
  • viral vectors and methods of the present invention are applicable to a variety of therapies of the hypothalamic-pituitary and neuroendocrine axis.
  • viral vectors can be used to introduce genes encoding proteins into pituitary cells that result in physiological modulation of hormone secretion from these cells. For example, by contacting pituitary cells with a viral vector containing the genetic information encoding somatostatin, growth hormone secretion from somatotroph adenomas can be modulated. In a significant proportion (>20%) of pituitary adenomas, both growth hormone and prolactin are secreted.
  • a vector containing, for example, both the somatostatin and tyrosine hydroxylase, or other combinations is especially useful in treating these plurihormonal tumors.
  • the following nonlimiting examples are provided to further illustrate the present invention.
  • Example 1 Vector construction and sequences The human tyrosine hydroxylase II gene is introduced via three viral vectors: a) modified human herpes simplex virus-1 vectors, pHSVth, containing a human tyrosine hydroxylase II gene under the control of the herpes immediate early 4/5 promoter (HSV EI-4/5); b) an adenovirus vector containing a human tyrosine hydroxylase gene; and c) an adeno-associated virus vector containing both a tyrosine hydroxylase and amino acid decarboxylase gene.
  • HSV EI-4/5 herpes immediate early 4/5 promoter
  • Human prolactinoma cultures are derived from human prolactinoma tissue obtained in the operating room from patients undergoing transphenoidal surgical resection of their tumors.
  • the tissue is kept in sterile saline on ice and transported immediately to the laboratory. There, the saline is carefully aspirated and the tissue laid out on a Petri Dish. Using two #11 scalpel blades heat sterilized previously, the tissue is minced carefully into small ribbons. Thereafter, a DNAase/trypsin solution in Hanks medium is added and the combined protease solution and prolactinoma sample are transferred using a 2 ml pipette into sterile capped Erlenmeyer flasks.
  • the tissue is incubated for 1-2 hours until the solution is cloudy, indicating complete digestion and the achievement of a dispersed cell state.
  • This solution is then spun on a table top centrifuge at 5,000 rpm for approximately 10 minutes, and the pellet is resuspended in 40% Dulbecco's modified Eagle's medium, 40% Ham's F12 medium, and 10% fetal calf serum, with 10,000 units of penicillin/streptomycin/100 ml of medium.
  • Cell counts are made using a hemacytometer and 10,000 to 100,000 cells are added to each well of a 48 well Nunclon dish pretreated with poly-1-lysine for 2 hours. The final number of cells per well in each dish is identical, but some variation in the number of cells between tumor samples does occur because of differing quantities of tumor specimen obtained.
  • Each data point represents the mean of at least 3 wells.
  • prolactin levels were determined in the media samples from the cultures treated with either controls or experimental viral vectors. The results for the levels of prolactin are demonstrated in Figures 5 and 6.

Landscapes

  • Life Sciences & Earth Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Health & Medical Sciences (AREA)
  • Genetics & Genomics (AREA)
  • Organic Chemistry (AREA)
  • Engineering & Computer Science (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Zoology (AREA)
  • Wood Science & Technology (AREA)
  • Molecular Biology (AREA)
  • Microbiology (AREA)
  • Biotechnology (AREA)
  • Biomedical Technology (AREA)
  • Biochemistry (AREA)
  • General Engineering & Computer Science (AREA)
  • General Health & Medical Sciences (AREA)
  • Medicinal Chemistry (AREA)
  • Medicines Containing Material From Animals Or Micro-Organisms (AREA)
  • Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)

Abstract

Des vecteurs viraux, qui contiennent des gènes sélectionnés codant des protéines, peuvent moduler la physiologie d'une tumeur pituitaire ou de l'hypophyse. Des procédés permettent d'utiliser de tels vecteurs dans le traitement des adénomes hypophysaires, des dysfonctionnements de l'axe hypothalame-hypophysaire et des maladies neuro-endocriniennes induites par l'hypophyse.
PCT/US1996/008191 1995-06-06 1996-05-31 Therapie genique de l'adenome hypophysaire et d'autres troubles endocriniens WO1996039416A1 (fr)

Priority Applications (1)

Application Number Priority Date Filing Date Title
AU60282/96A AU6028296A (en) 1995-06-06 1996-05-31 Gene therapy for pituitary adenoma and other endocrine disor ders

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
US47022795A 1995-06-06 1995-06-06
US08/470,227 1995-06-06

Publications (1)

Publication Number Publication Date
WO1996039416A1 true WO1996039416A1 (fr) 1996-12-12

Family

ID=23866760

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/US1996/008191 WO1996039416A1 (fr) 1995-06-06 1996-05-31 Therapie genique de l'adenome hypophysaire et d'autres troubles endocriniens

Country Status (3)

Country Link
AU (1) AU6028296A (fr)
CA (1) CA2221604A1 (fr)
WO (1) WO1996039416A1 (fr)

Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2022165313A1 (fr) 2021-02-01 2022-08-04 Regenxbio Inc. Thérapie génique de céroïdes-lipofuscinoses neuronales

Citations (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US5082670A (en) * 1988-12-15 1992-01-21 The Regents Of The University Of California Method of grafting genetically modified cells to treat defects, disease or damage or the central nervous system

Patent Citations (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US5082670A (en) * 1988-12-15 1992-01-21 The Regents Of The University Of California Method of grafting genetically modified cells to treat defects, disease or damage or the central nervous system

Non-Patent Citations (3)

* Cited by examiner, † Cited by third party
Title
EXPERIMENTAL NEUROLOGY, June 1994, Vol. 127, No. 2, ANTON et al., "Neural-Targeted Gene Therapy for Rodent and Primate Hemiprkinsonism", pages 207-218. *
JOURNAL OF NEUROCHEMISTRY, 1994, Vol. 62, No. 3, WU et al., "The Stability of Endogenous Tyrosine Hydroxylase Protein in PC-12 Cells Differs from that Expressed in Mouse Fibroblasts by Gene Transfer", pages 863-872. *
THE JOURNAL OF NEUROSCIENCE, December 1993, Vol. 13, No. 12, KANG et al., "Regulation of Dopamine Production by Genetically Modified Primary Fibroblasts", pages 5203-5211. *

Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2022165313A1 (fr) 2021-02-01 2022-08-04 Regenxbio Inc. Thérapie génique de céroïdes-lipofuscinoses neuronales

Also Published As

Publication number Publication date
CA2221604A1 (fr) 1996-12-12
AU6028296A (en) 1996-12-24

Similar Documents

Publication Publication Date Title
AU712775B2 (en) Adenoviral-vector-mediated gene transfer into medullary motor neurons
AU2009225335B2 (en) Methods and pharmaceutical compositions for healing wounds
KR102125658B1 (ko) 전립선 관련된 항원 및 백신 기재 면역치료 요법
US6685934B1 (en) Recombinant adenoviruses coding for basic fibroblast growth factors (BFGF)
US6884431B1 (en) Method for treating proliferative diseases with p27 and fusions thereof
JPH09507747A (ja) 中枢神経系の幹細胞の原位置修飾及び操作
EP0918867A2 (fr) Proteine-tyrosine-phosphatases de piegeage de substrats
JP2007524591A (ja) 同種拒絶反応の特異的阻害
Bankiewicz et al. Practical Aspects of the Development ofex Vivoandin VivoGene Therapy for Parkinson's Disease
IL173148A (en) Use of insulin for the preparation of pharmaceutical compositions for healing wounds
Joung et al. Effective gene transfer into regenerating sciatic nerves by adenoviral vectors: potentials for gene therapy of peripheral nerve injury
Rubenstein et al. Antisense oligonucleotide intralesional therapy for human PC‐3 prostate tumors carried in athymic nude mice
US20040224409A1 (en) Recombinant adenoviruses coding for brain-derived neurotrophic factor (BDNF)
Jeejeebhoy et al. Effects of hormones on the synthesis of α1 (acute-phase) glycoprotein in isolated rat hepatocytes
WO1996039416A1 (fr) Therapie genique de l'adenome hypophysaire et d'autres troubles endocriniens
WO1997039629A1 (fr) Vecteurs viraux incluant des polynucleotides codant des facteurs neurotrophiques ainsi que leur utilisation
Horellou et al. Adenovirus-mediated gene transfer to the central nervous system for Parkinson's disease
CA2118123A1 (fr) Methode visant a induire l'immunite tumorale
JP2001504123A (ja) Ｃｎｓ障害に対して保護するための成長因子を用いた予備治療
KR100403893B1 (ko) 글루타메이트 디 카르복실라제(gad) 활성을 암호화하는 재조합바이러스
US20020031493A1 (en) Recombinant adenoviruses coding for glial-derived cell neurotrophic factor (gdnf)
AU703793B2 (en) Recombinant adenoviruses coding for brain-derived neurotrophic factor (BDNF)
WO2001021217A2 (fr) Procede de traitement prophylactique ou therapeutique des cellules de la prostate
CN114729316A (zh) 用于治疗的产品和方法
HUP0104032A2 (hu) Új rendszer transzgén kifejeződés szabályozására

Legal Events

Date Code Title Description
AK Designated states

Kind code of ref document: A1

Designated state(s): AU CA JP

AL Designated countries for regional patents

Kind code of ref document: A1

Designated state(s): AT BE CH DE DK ES FI FR GB GR IE IT LU MC NL PT SE

121 Ep: the epo has been informed by wipo that ep was designated in this application
DFPE Request for preliminary examination filed prior to expiration of 19th month from priority date (pct application filed before 20040101)
ENP Entry into the national phase

Ref document number: 2221604

Country of ref document: CA

Ref country code: CA

Ref document number: 2221604

Kind code of ref document: A

Format of ref document f/p: F

122 Ep: pct application non-entry in european phase
点击 这是indexloc提供的php浏览器服务，不要输入任何密码和下载