US20120156170A1 - Drug composition for angiogenesis therapy - Google Patents

Drug composition for angiogenesis therapy Download PDF

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US20120156170A1
US20120156170A1 US13/393,062 US201013393062A US2012156170A1 US 20120156170 A1 US20120156170 A1 US 20120156170A1 US 201013393062 A US201013393062 A US 201013393062A US 2012156170 A1 US2012156170 A1 US 2012156170A1
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aav
angiogenesis
pgis
hpgis
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Tadashi Tanabe
Keiichi Hukuda
Takashi Kawakami
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    • A61K31/557—Eicosanoids, e.g. leukotrienes or prostaglandins
    • A61K31/558—Eicosanoids, e.g. leukotrienes or prostaglandins having heterocyclic rings containing oxygen as the only ring hetero atom, e.g. thromboxanes
    • A61K31/5585—Eicosanoids, e.g. leukotrienes or prostaglandins having heterocyclic rings containing oxygen as the only ring hetero atom, e.g. thromboxanes having five-membered rings containing oxygen as the only ring hetero atom, e.g. prostacyclin
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Definitions

  • the present invention relates to reinforcing materials for angiogenesis activities, which are useful pharmaceutical compositions for angiogenesis therapy
  • PAD peripheral arterial disease
  • arteriosclerosis brought by risk factors such as aging, smoking, diabetes, hypertension and metabolic disorders of lipids.
  • popularization of noninvasive examination tests for PAD detected a contract rate of about 2-3% below age of 50 years and that of about 20% over 75 years old. And approximately about 40% PAD patients are known to show no indications.
  • CLI critical limb ischemia
  • Bone marrow mononuclear cells and peripheral mononuclear cells are used for the cell therapy.
  • Recombinant angiogenesis factors are used for the gene therapy. These factors used for the gene therapy, for example, are vascular endotherial growth factor (VEGF), fibroblast growth factor (FGF), and hepatic growth factor (HGF). These genes for angiogenesis factors enhances angiopoiesis in ischemic tissues by expressing these factors. And by the use of these factors it was possibly ascertained that development of collateral blood vessels in model animals of heart or limb ischemia is promoted and/or enhanced (Circulation 90, 11-228-11-234 (1994).
  • VEGF vascular endotherial growth factor
  • FGF fibroblast growth factor
  • HGF hepatic growth factor
  • angiogenesis therapy Therapy for vasculopathy with above-mentioned angiogenesis factors is designated as “angiogenesis therapy”. More recently, using gene for above-mentioned angiogenesis factors therapeutic angiogenesis studies in ischemic and arterial diseases have been actively carried out.
  • a drug composition of angiogenesis therapy is one described in WO2002/000258 (patent reference 1).
  • the drug composition of angiogenesis therapy mentioned in patent reference 1 is the one containing gene for HGF or VEGF and prostacyclin synthase gene as effective components.
  • vascular endothelium In pathologic state vascular endothelium is injured and decreases in vasodialators such as nitric oxide, prostaglandin, endothelium-derived hyperpolarization factor (EDHF) and VEGF, and increases in vasoconstrictors, angoutensin-2, endothelin-1 etc. these vascular factors are can be useful for the treatment of PAD. Actually prostanoid is used for the clinical treatment of PAD and pulmonary arterial hypertention.
  • PGIS prostacyclin synthase
  • prostacyclin I 2 the synthase for prostacyclin
  • PGIS gene is a gene encoding an enzyme catalyzing the bio-synthesis of a remedy showing high effectiveness in treatment of experimental models of pulmonary hypertension and endoterium-injured arterioscrelosis.
  • the metabite shows activities inducing anti-platelet aggregation, vasodialation, and angiogenesis.
  • the gene therapy raises a long-term expression of the gene in local lesion, so that noxious side effects by the products always become issues.
  • Clinical research with growth factors is possibly accompanied by occurrence and development of tumors. In the clinical cases with PGI 2 drugs, however, there has been no report on tumorigenesis, and the safety of PGI 2 drugs is high because of their vasodialation and vaso-protective activities.
  • adeno-associated virus vector is judged to be the most suitable vector for the gene therapy due to the safe and active gene expression maintained for a relatively long-time.
  • plasmid or aveno virus are used as an expression vector. It is, however, with plasmid vectors that the strong and long gene expression is difficult to be obtained and that the frequent administration is necessary to maintain the expression level. Although a strong gene expression is obtained with the adenovirus vector, it has a short effect and the strong immunogenicity, and shows some concern about its safety. Conventionally, virus vectors have high transgenic activities, but their pathogenicities are often discussed in the human use. On the other hand AAV vectors deserve the attention because of a high efficiency and long-term expression in nondividing cells such as muscle and nerve cells, and their nonpathogenicity to human.
  • present inventors using AAV as a gene transfer vector, carried out the gene transfer in skeletal muscle of human prostacyclin synthase synthesizing prostaglandin I 2 (PG I 2 ) resulting in improvement of the condition of pulmonary hypertension model mouse.
  • PG I 2 prostaglandin I 2
  • intense expression of the transferred PGIS gene was recognized in the local skeletal muscle receiving the gene transfer. It is obtained from this experiment the information that the PGIS gene transfer is hopeful in treatment of limb ischemia.
  • the vector for gene therapy is expected to express long time in a possibly high level of the cure gene in the lesion.
  • many plasmids or non viral vectors were in the past gene therapy studies and with these vectors strong and long-term expression on the cure gene was difficult.
  • AVV adeno-associated virus
  • the present invention is concerned with the drug composition of angiogenesis used for the treatment or prevention of peripheral arterial disease.
  • This drug composition contains, as the active component, the adeno-associated virus (AVV) inserted with human gene for PGIS producing prostaglandin I 2 which at least induces the activities of anti-platelet aggregation, vasodialation, and/or angiogenesis.
  • AAV adeno-associated virus
  • this drug composition of angiogenesis therapy is characteristic of the recombinant adeno-associated virus (AVV) inserted with the gene for production of a factor inducing angiogenesis.
  • AAV adeno-associated virus
  • angiogenesis used for the treatment or prevention of peripheral arterial disease, which contains the adeno-associated virus (AVV) inserted with human gene for PGIS producing prostaglandin I 2 which at least induces the activities of anti-platelet aggregation, vasodialation, and/or angiogenesis or the recombinant adeno-associated virus (AVV) inserted with the gene for the angiogenesis factor.
  • AVV adeno-associated virus
  • AVV adeno-associated virus
  • the drug composition for angiogenesis therapy in the present invention includes a pharmaceutically permissible carrier such as the recombinant adeno-associated virus (AVV) inserted with the gene for human PGIS or human PGIS and the angiogenesis factor.
  • a pharmaceutically permissible carrier such as the recombinant adeno-associated virus (AVV) inserted with the gene for human PGIS or human PGIS and the angiogenesis factor.
  • the gene coding for the angiogenesis factor used in the drug composition for angiogenesis therapy of the present invention is possibly the entire or a part of the gene coding for a protein or a peptide, which can newly induces vasculogenesis.
  • VEGF vascular endotherial growth factor
  • the present invention presents a drug composition for angiogenesis therapy useful in treatment or prevention of peripheral arterial disease, which includes human gene for PGIS (hPGIS) producing prostaglandin I 2 which induces the activities of vasodialation, and/or anti-platelet aggregation, and the recombinant adeno-associated virus (AVV) inserted with the gene for the angiogenesis factor
  • hPGIS human gene for PGIS
  • AAV adeno-associated virus
  • the “gene for angiogenesis” inserted in adeno-associated virus (AAV) is, similarly to the human PGIS gene, the gene coding for the entire or a part of a protein or a peptide, which newly induces vasculogenesis and shows either activity of inhibition of platelet aggregation, vasodialation, angiogenesis.
  • VEGF endotherial growth factor
  • VEGF-2 acidic FGF
  • bFGF basic FGF
  • TGF- ⁇ TGF- ⁇
  • PD-ECGF platelet-derived endotherial growth factor
  • TGF- ⁇ tumor necrosis factor- ⁇
  • insulin-like growth factor angiopoieti-1 etc.
  • HIF-1 and ets family translation factors including ets-1 which regulate the gene expression of VEGF.
  • VEGF gene in the present invention indicates the gene coding for VEGF protein and, in some case, simply designates the expression vector (? plasmid) encoding the VEGF gene.
  • a cDNA for VEGF is inserted into the virus vector mentioned after.
  • human VEGF four subtypes (VEGF121, VEGF165, VEGF189, VEGF206) formed by alternative splicing in transcription of the human gene have been reported (Science, 219, 983, (1983); J. Clin. Invest., 84, 1470 (1989); Biochem. Biophys. Res. Commun, 161, 851 (1989)).
  • the gene for VEGF165 with the biologically strongest activity is preferable.
  • the VEGF gene subjected to modifications are classified as the VGEF gene as far as they express the active protein with an angiogenesis activity.
  • VGF165 The gene for VGF165 is easily cloned or modified by a man of skill in related fields according to information on the sequence registered in the literature (i.e. Science, 246, 1306 (1989)) or data bases.
  • VEGF vascular endotherial growth factor
  • Previously described gene for angiogenesis factors can be used singly or in combination with multiple gene.
  • PGI 2 produced by hPGIS shows the activities of vasodialation, accentuation of vasopermiability and anti-platelet aggregation and a combination of gene for PGIS and an angiogenesis factor brings a more remarkable combination effect.
  • the material having activities of vasodialation and/or anti-platelet aggregation is defined as the compound which produces PGI 2 showing activities of vasodialation and/or anti-platelet aggregation, or enhances the production of PGI 2 . Properly it indicates the compound which enhances synthesis, production of induction of prostaglandin (I 2 ?).
  • the suitable compound can be any material including gene, proteins, low molecular weight chemicals etc.
  • the gene for PGIS is utilized as the compound induces the enzyme synthesizing PGI 2 .
  • the nucleotide sequence of the gene for PGIS are registered in public data bases and its gene is easily cloned by a man of skill in related fields.
  • PGIS gene indicates the gene coding for PGIS.
  • PGIS gene constructed into an expression plasmid is also described as “PGIS gene”.
  • PGIS gene is the clone obtained by inserting the PGIS cDNA recorded in B.B.R.C., Vol. 200, No. 3, p 1728-1734 (1994) and WO95/30013 into a proper expression vector.
  • the expression constructs with modified PGIS gene are included in the PGIS gene as far as they are the gene coding for protein with the function of PGIS in the present investigation.
  • the present PGIS gene, as well as the VEGF gene, is also easily cloned on the basis of the sequence information described in the previously described reference and registered in public data bases by a man of skill in related fields and its modification of the clone is easily carried out.
  • the expected activity of the protein coded by the present gene can be examined by assaying products, for example, using an enzyme immunoassay using 6-keto Prostaglandin F1 ⁇ enzyme immunoassay kit (Cayman, Co., catalog #515211), or thin-layer chromatography detecting the products of prostacyclin synthase.
  • PGI 2 as the compound with activities of vasodialation and/or anti-platelet aggregation or PGI 2 -inducing materials can be singly used or in combination of multiple compounds.
  • the objective gene is inserted into the adeno-associated virus (AVV), and can be transfered into the target cells by infection of the recombinant virus (AVV) constructed by insertion of the gene into the adeno-associated virus (AVV) as a vector.
  • AVV recombinant virus
  • types 1, 2, 5, 8 of adeno-associated virus (AVV) are suitable, but types 1 or 2 are preferred.
  • the vector is used for the purpose of transfer of the gene into the target region of the lesion.
  • This selection of adeno-associated virus (AVV) as a vector is due to the efficient transfer of gene into non-dividing cells such as muscle, nerve and liver cells etc. and to the possible long-term expression maintained in these target cells.
  • AMV adeno-associated virus
  • the gene therapy drug in the present invention consisting of drug composition of angiogenesis is preferred to be type 1 or 2 of adeno-associated virus (AVV) with an efficient transfer activity of the gene into non-dividing cells such as muscle, nerve and liver cells etc.
  • AAV adeno-associated virus
  • the transfer method of the gene-therapeutic drug to the patient there are two methods: the in vivo method where the gene-therapeutic drug is directly delivered into the body; the ex vivo method where it is ex vivo transfected into a specified cells isolated from the human body and returned to the body (Nikkei Science (in Japanese), April issue, 1994, pp. 20-45; Gekkanyakuji (in Japanese), 36(1), 23-48, 1994; Separate Volume Jikken-igaku, 12(15), 1994; Handbook for R&D of Gene Therapy (in Japanese), ed. The Japan Society of Gene Therapy, NTS Co., 1999).
  • the in vivo method is preferred in the present invention.
  • the drug for gene therapy When the drug for gene therapy is given to the patient, it is delivered in the pathway dependent on the disease as the therapeutic target and the target organ etc.
  • the drug for gene therapy is injected into veins, arteries, hypodermic or intracutaneous tissues, and muscles, or directly given to the tissues with the lesion.
  • the injection as the dose form containing the active ingredient of a gene can be prepared by common methods.
  • the injection is possibly prepared as follows: the ingredient, for example, is dissolved in a suitable solvent (a buffer such as PBS, saline, sterilized water etc) if necessary, subjected to mechanical sterilization by filtration, and filled in sterilized vessels.
  • a suitable solvent a buffer such as PBS, saline, sterilized water etc
  • a commonly used carrier can be added to the injection.
  • the injection in use of a liposome such as the HVJ-liposome, the injection can be a suspension, a frozen form or a frozen drug after centrifugal concentration.
  • the drug can be embedded around the lesion or slowly and continuously delivered with an osmotic pump.
  • the content of the active pharmaceutical ingredient (API) of the gene can be adjusted according to the disease as the treatment aim, age of patient, body weight etc., is generally in a range of 0.001-100 mg, preferably 0.01-10 mg, which is suitably given to a patient once per days or months.
  • anti-platelet drug drugs with activities of vasodialation and/or anti-platelet aggregation
  • their administration methods and dose can be determined according to their protocols.
  • the dose form and administration method are as follows.
  • the drug when the drug is orally administered, it is done in the dose form usually used in the specified field.
  • the drug When the drug is parenterally dispensed, it is administered as the dose form of a focal administration drug, an intrarectal administration drug, an injection, a nasogastric drug etc.
  • the dose form of capsules, tablets, pills, powder, drop, suppository, liquid formulation etc. is conceived.
  • a sterilized solution, suspension, and emulsion etc, such as water, a water-propylene glycol mixed solution, a buffered solution, 0.4% saline etc. are conceived as the injection form.
  • the dose form of cream, ointment, lotion, transdermal absorption agent etc. can be chosen as a focal administration drug.
  • compositions with pharmaceutically tolerably excipients and additives are prepared to the formulations with pharmaceutically tolerably excipients and additives, by common methods employed in the pharmaceutical field.
  • Pharmaceutically tolerably excipients and additives are carriers, binders, incenses, buffering agents, thickening agents, tinctions, stabilizers, emulsifiers, dispersing agents, suspensions, antiseptics, pH-adjusting agents, tonicity-adjusting agents infiltrate agents etc.
  • pharmaceutically tolerably carriers for example, are magnesium carbonate, lactose, pectin, starch, methylcellulose etc.
  • the drug composition When the previously described drug composition is given to the patient, it can be delivered in the pathway dependent on the disease as the therapeutic target and the target organ etc. For example it may be injected into veins, arteries, hypodermic or intracutaneous tissues, and muscles, or directly given to the tissues with the lesion. Additionally the drug composition is also possible to be administered orally or in its suppository form. Incidentally dose and administration frequency of the drug composition is depends on the symptom, age, body weight of a patient, and the administration form. Generally the dose per an adult is in a range of 0.001-100 mg a day, preferably 0.01-10 mg, which is suitably given to a patient once or multiple times divided.
  • compositions with a low-molecular weight chemical or protein as an active ingredient can be administered together with the gene-therapy drug containing the gene for angiogenesis, or separately at regular intervals.
  • the drug composition of angiogenesis therapy in the present investigation is described above.
  • the present drug composition is useful if treatment or prevention of the peripheral arterial disease, which needs the angiogenesis therapy.
  • FIG. 1 shows subcutaneous blood perfusion by the laser Doppler method of ischemic limbs treated with adeno-associated virus-mediated PGIS gene (AAV-PGIS group), saline (control group), adeno-associated virus-mediated EGFP gene (AAV-EGFP (enhanced green fluorescence protein) group) and the corresponding macroscopic images.
  • AAV-PGIS group adeno-associated virus-mediated PGIS gene
  • saline control group
  • AAV-EGFP enhanced green fluorescence protein
  • FIG. 3 indicates the improvement in limb necrosis rate of AAV-PGIS group mice in comparison with those of control and AAV-EGFP group mice.
  • GPDH mouse glyceraldehydes-3-phosphate dehydrogenase
  • FIG. 5 shows the western blot analysis of the expression of human PGIS (hPGIS) in ischemic muscles received an injection of control (saline), AAV-EGFP or AAV-PGIS.
  • FIG. 6 shows immunostaining images of human PGIS (A) and mouse von Willebrand factor (vWF) (B), and relative capillary vessel area in hind limb muscles received an injection of control (saline), AAV-EGFP or AAV-PGIS.
  • A Human PGIS expressed in hind limb muscles received an injection of control (saline), AAV-EGFP or AAV-PGIS was immunostained by anti-human PGIS antibody.
  • B Mouse vWF as a neovascularization marker expressed in hind limb muscles in the three groups was immunostained by Anti-vWF antibody.
  • C vWF positive area of the images in FIG. 6B was quantitatively analyzed.
  • the expression vector for human PGIS was constructed as described previously.
  • the blunted HindIII/BamHI fragment of the full-length human PGIS cDNA was ligated into the blunted XhoI site of the pUC-CAGGS expression plasmid.
  • pUC/PGIS was transfected into NIH3T3 cells, and conversion of [ 14 C]-PGH 2 to 6-keto-[ 14 C]PGF1 ⁇ was measured.
  • pUC-CAGGS vector lacking the insert served as the control vector.
  • Human PGIS genes were then inserted into AAV-CAG plasmids, and AAV-hPGIS vectors were constructed.
  • AAV-EGFP (enhanced green fluorescent protein) vector for control experiments was also prepared as described previously.
  • mice were divided into 3 groups (control, AAV-EGFP and AAV-hPGIS).
  • the AAV-hPGIS group was administered AAV type1-hPGIS (1.0 ⁇ 10 11 ) into left thigh muscle.
  • the control group was administered the vehicle (0.9% saline) into the left thigh muscle.
  • As negative control the AAV-EGFP group was administered AAV type1-EGFP (1.0 ⁇ 10 11 ) into same site.
  • the left femoral artery of the mice was ligated to produce hind limb ischemia model.
  • the incidence of limb necrosis was evaluated and blood perfusion rate of the mice was measured using Laser Doppler perfusion imaging (LDPI) system.
  • LDPI Laser Doppler perfusion imaging
  • mice in the groups of hPGIS, control, and AAV-EGFP were administered AAV type1-hPGIS (1.0 ⁇ 10 11 , v.g./body), into left thigh muscle of C57/BALB6 mice (8 week-old)
  • mice The three groups (AAV-hPGIS, CONTROL and AAV-EGFP) of mice are subjected to ischemia in left hind limbs, and the blood perfusion rate in the ischemic left (L) and normal right (R) hind limb was measured with a Laser Doppler perfusion imaging (LDPI) system (Moor LDI, Moor Instruments, USA). The measurement was carried out by taking LDPI of perfusion state of ischemic hind limbs in each group. The results are shown in FIG. 1 .
  • LDPI Laser Doppler perfusion imaging
  • FIG. 1 optical (Macro) and the corresponding Laser Doppler (LDP) perfusion images obtained by measurement with LDPI system.
  • LDP Laser Doppler
  • FIG. 1 the images obtained by LDPI method are represented in color.
  • the index for color images representing perfusion state shown below. The index from left to right changes from dark blue to red. Low or nil blood perfusion was displayed as dark blue, and the highest perfusion intervals were displayed as red up to white (maximum).
  • RNA extraction and quantitative RT-PCR were performed as described previously. Quantitative RT-PCR was performed with TaqMan probes (Applied Biosystems): human PGIS (Hs00168766_m1), mouse PGIS (Mm00447271_m1), mouse VEGF (Mm01281449_m1), FLK-1 (Mm01222419_m1), FLT-1 (Mm00438971m1), and Rodent GAPDH.
  • the mRNA levels were normalized by comparison to GAPDH mRNA. In FIG. 4 data are mean ⁇ SD. *P ⁇ 0.05; ns, not significant.
  • Membrane fractions of skeletal muscle were prepared by homogenization of thigh muscle in ice-cold buffer as described Immunodetection was performed on membrane extracts with an antibody to human PGIS. The results are shown in FIG. 5 .
  • FIG. 6(A) illustrates microscopic images showing expression of hPGIS in left hind limb muscles in groups of AAV-hPGIS, Control and AAV-EGFR
  • the each muscle in (A) was stained by immnohistochemical methods.
  • B Typical examples of microscopic images of von Willebrand factor (vWF)-stained left hind limb muscles mouse.
  • C The results of quantitative analysis of microvessels in left hind limb muscles subjected to immunofluorecence staining of vWF. In FIG. 6C data are mean ⁇ SD. *P ⁇ 0.05; ns, not significant.
  • mice 8-weeks control mice (C57/BL/6). Mice were administered AAV type1-hPGIS vector into the left thigh muscle. For analysis the left thigh muscle from mice treated with before and after 2, 4, 8, 12 weeks vector administration. The results, as analyzed by quantitative RT-PCR, indicates a strong human PGIS mRNA expression from 2 weeks after gene transfer. The strong expression was maintained even after 12 weeks. The results are shown in Table 1.
  • mice were administered AAV1-human PGIS vector (1.0 ⁇ 10 11 (v.g./body)).
  • LDP laser Doppler blood perfusion
  • mRNA in ischemic limb tissues was analyzed using quantitative RT-PCR. Compared with Control and AAV-EGFP groups, a remarkably significant expression of human PGIS gene was observed in AAV-hPGIS group (see FIG. 4A ) and the expression of intrinsic mouse PGIS gene was not significantly different among the three group mice. Furthermore, expression of murine mRNA for VEGF, FLK-1 and FLT-1 was analyzed to study neovascularization in the ischemic limb tissues. Consequently, it was significant that expression of VEGF mRNA in AAV-PGIS group was twice compared with that in the other two groups (See FIG. 4C ).
  • vVW-positive microvessels was quantitatively analyzed using Image J software. The capillary density was expressed as the ratio between the total area of capillary vessels and the total skeletal muscle area.
  • neovascularization at the capillary level occurred in the ischemic skeletal muscles administered AAV-PGIS, suggesting reduction of ischemic injury of skeletal muscles.
  • AAV type 1 is known to have an ability of the strongest and long-term expression in skeletal muscles. It is reported by the present inventor et al that AAV type 1-human PGIS (AAV-hPGIS) shows a long-term strong gene transfer efficiency in the in vitro and in vivo control experiments. Additionally gene expression of AAV type 1 is limited in the local site of administration, and it is not found in remote organs. This characteristic of the expression can reduce the side effect of the gene therapy in remote organs, and is useful in the treatment of patients suffering from serious limb ischemia, which are complicated with arteriosclerosis-induced diseases in other organs.
  • skeletal muscles was subjected to immnostaining with anti-vWF antibody in order to examine whether neovascularization occurs or not.
  • AAV-PGIS group neovascularization was confirmed at the capillary level.
  • EVG-staining for elastic fibers in skeletal muscles was carried out and significant changes were not observed in all treated groups, suggesting that the neovascularization at capillary level occurs in skeletal muscles administered AAV-hPGIS.

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* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP1308517A1 (en) * 2001-10-31 2003-05-07 Aventis Pharmacueticals Products Inc. Vectors for expressing multiple transgenes
US20060099183A1 (en) * 1999-08-02 2006-05-11 Pierre Zoldhelyi Use of recombinant adeno-associated virus vector (rAAV) for the prevention of smooth muscle cell proliferation in a vascular graft
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* Cited by examiner, † Cited by third party
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DE69535091T2 (de) 1994-04-28 2007-01-04 Tanabe, Tadashi, Toyonaka Humane prostacyclinsynthase
CA2230819C (en) 1995-08-29 2009-04-14 Sumitomo Pharmaceuticals Co., Ltd. Medicament comprising hgf gene
WO2002000258A1 (en) * 2000-06-27 2002-01-03 Anges Mg, Inc. Medicinal compositions for angiogenic therapy

Patent Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US20060099183A1 (en) * 1999-08-02 2006-05-11 Pierre Zoldhelyi Use of recombinant adeno-associated virus vector (rAAV) for the prevention of smooth muscle cell proliferation in a vascular graft
US7276490B1 (en) * 2000-05-22 2007-10-02 Tadashi Tanabe Medicinal compositions containing prostacylin synthase gene
EP1308517A1 (en) * 2001-10-31 2003-05-07 Aventis Pharmacueticals Products Inc. Vectors for expressing multiple transgenes

Non-Patent Citations (7)

* Cited by examiner, † Cited by third party
Title
Authors unknown, downloaded 5/2/17, https://en.wikipedia.org/wiki/P-value, (no journal/volume/issue/pages), as a PDF, 9 pages long. *
Fu, et al. (2005) "Optimization of AAV Serotypes and Promoter for Increased Distribution of Expression in Both Somatic and Central Nervous System of Mice", Molecular Therapy, 11(Supp 1): S332. *
Gupta, et al. (2003) "Cyclooxygenase-1 is Overexpressed and Promotes Angiogenic Growth Factor Production in Ovarian Cancer", Cancer Research, 63: 906-11). It is noted that several of the other proteins to be encoded also promote angiogenesis, e.g., E2F-1, ENOS. *
Haidet (2008) "Long-term enhancement of skeletal muscle mass and strength by single gene administration of myostatin inhibitors", Proceedings of the National Academy of Science, USA., 105(11): 4318-22. *
Hiraoka, et al. (2003) "Enhanced Therapeutic Angiogenesis by Cotransfection of Prostacyclin Synthase Gene or Optimization of Intramuscular Injection of Plasmid DNA", Circulation, 108: 2689-96 *
Kawakami, et al. (2007) "AAV-PGIS gene transfer improves hypoxia-induced pulmonary hypertension in mice", Biochemical and Biophysical Research Communications, 363(3): 656-61. *
Yan, et al. (2005) "Superior neovascularization and muscle regeneration in ischemic skeletal muscles following VEGF gene transfer by rAAV1 pseudotyped vectors", Biochemical and Biophysical Research Communications, 336: 287-98. *

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