Pharm/Biotech
Resources

Outsourcing Guide

Cont. Education

Software/Reports

Training Courses

Web Seminars

Jobs

Buyer's Guide

Home Page

Pharm Patents /
Licensing

Pharm News

Federal Register

Pharm Stocks

FDA Links

FDA Warning Letters

FDA Doc/cGMP

Pharm/Biotech Events

Consultants

Advertiser Info

Newsletter Subscription

Web Links

Suggestions

Site Map
 

 

 

 

Title:  Medicament comprising HGF gene

United States Patent:  6,248,722

Inventors:  Morishita; Ryuichi (Osaka, JP); Ogihara; Toshio (Mino, JP); Nakamura; Toshikazu (Takatsuki, JP); Tomita; Tetsuya (Toyonaka, JP); Ochi; Takahiro (Kobe, JP)

Assignee:  Sumitomo Pharmaceuticals Company, Limited (Osaka, JP)

Appl. No.:  029497

Filed:  June 9, 1998

PCT Filed:  August 22, 1996

PCT NO:  PCT/JP96/02359

371 Date:  June 9, 1998

102(e) Date:  June 9, 1998

PCT PUB.NO.:  WO97/07824

PCT PUB. Date:  June 3, 1997

Foreign Application Priority Data:  Aug 29, 1995[JP] (7-245475); Feb 20, 1996[JP] (8-058467)

Abstract

The present invention relates to a medicament comprising a HGF gene. The medicament of the present invention may be topically applied to the target organs so that the effects can be selectively exhibited, resulting in minimizing the side effects of HGF.

BEST MODE FOR CARRYING OUT THE INVENTION

The "HGF gene" employed in the present invention indicates a gene capable of expressing HGF. Thus, so long as a polypeptide expressed has substantially the same effect as that of HGF, the HGF gene may have a partial deletion, substitution or insertion of the nucleotide sequence, or may have other nucleotide sequence ligated therewith at the 5'-terminus and/or 3'terminus thereof. Typical examples of such HGF genes include HGF genes as described in Nature, 342, 440 (1989), Japanese Patent KOKAI (Laid-Open) No. 5-111383, Biohem. Biophys. Res. Commun., 163, 967 (1989), etc. These genes may be used in the present invention.

The HGF gene is incorporated into an appropriate vector and the HGF gene-bearing vector is provided for use. For example, the HGF gene may be used in the form of a viral vector having the HGF gene as described hereinafter, or in the form of an appropriate expression vector having the HGF gene.

The "pharmaceutical composition" used in the present invention indicates a medicament for the treatment or prevention of human diseases, which is attributed to the pharmacological activities of HGF. For example, exemplified are medicaments for the treatment or prevention of the diseases given hereinabove.

According to the present invention, the HGF gene is introduced into cells wherein HGF is expressed in those cells to exhibit the pharmacological actions. Thus, the medicament of the present invention is effectively applicable to the diseases for which HGF itself is effective.

Where the HGF gene is introduced into, e.g., cells, the growth of vascular endothelial cells is accelerated, while undesired growth of vascular smooth muscle cells is not accelerated, as demonstrated in the Examples hereinafter. Moreover, as demonstrated in the Example hereinafter, where the HGF gene is introduced into the heart in vivo animal test using rats, angiogenesis is observed. Therefore, the HGF gene is effective for the treatment and prevention of arterial disorders, in particular, various diseases caused by disturbance which mainly involves abnormal proliferation of vascular smooth muscle cells (e.g., restenosis after percutaneous transluminal coronary angioplasty (PTCA), arteriosclerosis, insufficiency of peripheral circulation, etc.), and for the treatment and prevention of diseases such as myocardial infarction, myocardia, peripheral angiostenosis, cardiac insufficiency, etc. HGF itself is also useful for the treatment and prevention of the diseases as described above, since HGF promotes the proliferation of vascular endothelial cells but does not promote the growth of vascular smooth muscle cells. The pharmacological effects of the HGF gene are attributed to those of HGF itself.

As demonstrated in the Examples hereinafter, introduction of the HGF gene into the joint results in promoting repair of articular cartilage cells thereby to promote the proliferation of proteoglycan-synthesizing cells. Therefore, the HGF gene is effective for the prevention and treatment of various cartilage injuries such as osteogenetic abnormality, arthritis deformans, discopathy deformans, fracture repair and restoration insufficiency, trauma caused by sporting, key puncher's disease, etc. HGF itself is useful for the treatment and prevention of the diseases described above, since HGF promotes repair and growth of cartilage cells. The effects of the HGF gene are based on those of HGF itself.

"Liposome" is a closed vesicle of lipid bilayer encapsulating an aqueous compartment therein. It is known that the lipid bilayer membrane structure is extremely similar to biological membranes. To prepare the liposomes of the present invention, phospholipids are employed. Typical examples of phospholipids are phosphatidylcholines such as lecithin, lysolecithin, etc.; acidic phospholipids such as phosphatidylserine, phosphatidylglycerol, phosphatidylinositol, phosphatidylic acid, etc.; or phospholipids obtained by replacing an acyl group(s) of these acidic phospholipids with lauroyl, myristoyl, oleoyl, etc.; and sphingophospholipids such as phosphatidylethanolamine, sphingomyelin, etc. Neutral lipids such as cholesterol may also be added to these phospholipids. The liposomes may be prepared, in a conventional manner, from naturally occurring materials such as lipids in normal cell membranes. The liposomes containing the HGF gene of the present invention may be prepared, for example, by suspending a thin layer of purified phospholipids in a solution containing the HGF gene and then treating the suspension in a conventional manner such as ultrasonication.

The liposomes containing the HGF gene of the present invention may be appropriately fused to viruses, etc. to form membrane fusion liposomes. In this case, it is preferred to inactivate viruses, e.g., through ultraviolet irradiation, etc. A particularly preferred example of the membrane fusion liposome is a membrane fusion liposome fused with Sendai virus (hemagglutinating virus of Japan: HVJ). The membrane fusion liposome may be produced by the methods as described in NIKKEI Science, April, 1994, pages 32-38; J. Biol. Chem., 266 (6), 3361-3364 (1991), etc. In more detail, the HVJ-fused liposome (HVJ-liposome) may be prepared, e.g., by mixing purified HVJ inactivated by ultraviolet irradiation, etc. with a liposome suspension containing the HGF gene vector, gently agitating the mixture and then removing unbound HVJ by sucrose density gradient centrifugation. The liposomes may be bound to substances having an affinity to target cells, thereby to enhance an efficiency of gene introduction into the target cells. Examples of the substances having an affinity to the target cells include ligands such as an antibody, a receptor, etc.

For introduction of the HGF gene into cells, conventional methods are employed, which are roughly classified into introduction via viral vectors and other strategies (NIKKEI Science, April, 1994, pages 20-45; GEKKAN YAKUJI, 36 (1), 23-48 (1994) and references cited therein). Both methods are available for the preparation of the medicament of the present invention.

The former method using viral vectors comprises the step of incorporating the HGF gene into, e.g., a retrovirus, an adenovirus, an adeno-related virus, a herpes virus, a vaccinia virus, a poliovirus, a sindbis virus or other RNA viruses. Of these viruses, a retrovirus, an adenovirus and an adeno-related virus are particularly preferably employed for the introduction.

Examples of the other methods include the liposome method, lipofectin method, microinjection method, calcium phosphate method, electroporation method. Of these methods, particularly preferred is the liposome method.

For practical use of the HGF gene as a medicament, it is advantageous to introduce the HGF directly into the body (in vivo method). Alternatively, certain cells are collected from human, the HGF gene is then introduced into the cells outside the body and the HGF gene-introduced cells are returned to the body (ex vivo method). These methods are described in NIKKEI Science, April, 1994, pages 20-45; GEKKAN-YAKUJI, 36 (1), 23-48 (1994) and references cited therein. Any of these methods are suitably chosen depending upon a disease to be treated, target organs, etc. and applied to the medicament compositions of the present invention.

The in vivo method is less costly, less laborious and therefore more convenient than the ex vivo method, but the latter method provides a higher efficiency of introduction of the HGF gene into cells.

Where the medicament of the present invention is administered by the in vivo method, the medicament may be administered through any route appropriate for diseases to be treated, target organs, etc. The medicament may be administered intravenously, intraarterially, subcutaneously, intramuscularly, etc., or directly to the objective organ of diseases, e.g., kidney, liver, lung, brain, nerve, etc. Direct administration to the objective site can treat the target organ selectively. For example, in gene therapy using a gene for restenosis after PTCA, the composition may be administered intraarterially (JIKKEN-IGAKU, 12 (extra issue 15), 1298-1933 (1994). Preferably, the medicament of the present invention is applied at the tip of a balloon used for PTCA and rubbed the tip against blood vessel, whereby the medicament may be introduced directly into vascular endothelial cells and vascular smooth muscle cells.

Where the ex vivo method as described above is used to introduce the HGF gene, human cells (e.g., lymphocytes or hematopoietic stem cells) are harvested in a conventional manner and the harvested cells are sensitized with the medicament of the present invention for gene introduction. Thereafter the HGF-producing cells are inserted back to human.

Where the medicament is administered by the in vivo method, the medicament may take various preparation forms, including the form of liquid preparation. In general, the medicament may be preferably prepared into an injection comprising the HGF gene as an active ingredient. If necessary and desired, conventional carriers may be added to the composition. The injection may be prepared in a conventional manner, e.g., by dissolving the HGF gene in an appropriate solvent (e.g., sterilized water, a buffered solution, a physiological saline solution, etc.), filtering the solution through a filter, etc. for sterilization, filling up the solution in a sterile container. The medicament may be prepared using the HGF gene-incorporated viral vector, instead of the HGF gene itself. Where the liposomes containing the HGF gene embedded therein (or HVJ-liposomes) are employed, the medicament may be in the form of liposome preparations such as a suspension, a frozen preparation, a centrifugally concentrated frozen preparation, etc.

The content of the HGF gene in the medicament may be appropriately varied depending upon diseases to be treated, target organs, patients' ages or body weights, etc. However, it is appropriate to administer in a dose of 0.0001 mg to 100 mg, preferably 0.001 mg to 10 mg when calculated as the HGF gene. The dose may be divided into several days or a few months.

Claim 1 of 6 Claims

What is claimed is:

1. A method for treating a disease in a subject for which HGF is effective, comprising administering intramuscularly to the subject an expression vector containing a HGF gene in a therapeutically effective amount.

____________________________________________
If you want to learn more about this patent, please go directly to the U.S. Patent and Trademark Office Web site to access the full patent.

 

 

[ Outsourcing Guide ] [ Cont. Education ] [ Software/Reports ] [ Training Courses ]
[ Web Seminars ] [ Jobs ] [ Consultants ] [ Buyer's Guide ] [ Advertiser Info ]

[ Home ] [ Pharm Patents / Licensing ] [ Pharm News ] [ Federal Register ]
[ Pharm Stocks ] [ FDA Links ] [ FDA Warning Letters ] [ FDA Doc/cGMP ]
[ Pharm/Biotech Events ] [ Newsletter Subscription ] [ Web Links ] [ Suggestions ]
[ Site Map ]