Drug – bio-affecting and body treating compositions – Designated organic active ingredient containing – Peptide containing doai
Reexamination Certificate
1999-04-12
2004-09-07
Housel, James (Department: 1648)
Drug, bio-affecting and body treating compositions
Designated organic active ingredient containing
Peptide containing doai
C514S002600, C514S011400, C514S016700, C530S300000
Reexamination Certificate
active
06787520
ABSTRACT:
The invention relates to an extraordinarily efficient method of inactivating lipid-enveloped viruses such as herpes or retroviruses in biological or biotechnological—particularly pharmaceutical—products, as well as in cell cultures by adding a cyclic lipopeptide or a mixture of lipopeptides or salts or esters thereof at specific concentrations. Lipopeptides were found to have a surprisingly high inactivation potential for lipid-enveloped viruses and in addition, they offer the advantage of an exceedingly low in vivo toxicity, so that the step of removing the inactivating agent from pharmaceutical products or cell cultures can be omitted. The invention is also directed to new antiviral lipopeptides which belong to the surfactins.
At the latest, the AIDS epidemic has brought the realization into the mind of the general public that not only HI viruses but rather, a variety of agents pathogenic for humans may be transmitted, for example, via blood transfusions, pharmaceuticals, transplantations, etc.. In general, it is obligatory today that each pharmaceutical agent prepared from biological material or having come in contact with same is rated as potentially contaminated by microbes or viruses, and that infection-related safety is verified. As a result of the development of molecular-biological methods for manufacturing pharmaceutical agents, the risk of infection by various microbial contaminants has increased further. Animal or human cell lines are frequently used in the biotechnological production of pharmaceuticals. In these cells, in particular, virus infections by endogenous viruses, latent virus infections, or contaminations cannot be excluded completely. The infection-related safety of biotechnological pharmaceuticals, e.g., vaccines, monoclonal antibodies, hormones, or recombinant proteins therefore necessitates removal of any infectious, undesirable particles, which, in principle, may be associated with a considerable loss of research time and means, or productivity. Virus safety of blood and blood products can only be guaranteed by testing and selecting the blood donations, in combination with evaluating and prophylactically using efficient and reliable methods of virus inactivation and elimination.
To inactivate and eliminate viruses from pharmaceutical products, various methods are employed alone or in combination. Chromatographic methods, pH shift, extraction, and fractionation using various organic solvents, salt precipitation, heat treatment, and filtration techniques are employed in case of structurally simple and stable products [Rabenau & Doerr (1990), “Die Infektionssicherheit biotechnologischer Pharmazeutika aus virologischer Sicht”, p. 58, GIT VERLAG GmbH, Darmstadt]. Where sensitive or complex biological materials are involved, substances having an antiviral effect are frequently used. Inter alia, the following methods are employed:
the combined use of solvents (e.g., extraction with ether) and synthetic detergents (e.g., Triton X-100) [B. Horowitz et al. (1985), Transfusion 25, 516-522];
the use of &bgr;-propiolactone in combination with UV light, as well as methylene blue in combination with photoactivation [W. Stephan (1989) pp. 122-127, in J.-J. Morgenthaler (Ed.); Virus Inactivation in Plasma Products; Curr. Stud. Hematol. Blood Transfus. No. 56, Karger, Basel];
pasteurization of liquid material [T. Nowak (1992), Biologicals 20, 83-85];
heating of lyophilized material [D. Piszkiewicz et al. (1989), pp. 44-54, in J.-J. Morgenthaler (Ed.), Virus Inactivation in Plasma Products., Curr. Stud. Hematol. Blood Transfus. No. 56, Karger, Basel];
irradiation with gamma rays (e.g., cobalt-60) [B. Horowitz et al. (1988), Transfusion 25, 523-527].
The literature describes a number of virus inactivation methods for blood products, particularly human blood plasma. Thus, in U.S. Pat. No. 4,591,505, A. M. Prince discloses an inactivation method for hepatitis B virus wherein the blood products are added with alcohol and either a non-ionic detergent or an ether or a mixture of both as virus-inactivating agent. Polyoxyethylene derivatives or sulfobetains are used as non-ionic detergents.
In U.S. Pat. No. 4,841,023 and in Vox-Sang. 54, 14-20 (1988), S. Karger A G, Basel, B. Horowitz describes the inactivation of lipid-containing viruses in blood products by fatty acids, and in U.S. Pat. No. 4,613,501 by C
1
-C
4
alkyl oleic acid.
In EP 0,050,061 E. Shanbrom discloses a method of reducing undesirable effects such as pyrogenicity, hepatitis infectiousness, and aggregation in biological and pharmaceutical products, particularly blood products as well, using a treatment with non-denaturing amphiphilics such as non-ionic surfactants (e.g., Tween 80).
The inactivation and elimination of viruses from cell cultures is effected using antiviral substances which normally inhibit virus replication.
None of the inactivation methods used up to now can safely inactivate or eliminate all viruses which may occur in biological material. Methods such as pasteurization or heat treatment normally require the use of stabilizers, and in addition, there is the problem of protein denaturation. To date, the use of solvents and synthetic surfactants considered as suitable for inactivating lipid-enveloped viruses could not be assessed as entirely safe due to deviating results in inactivation kinetics or lacking systematic investigations as a result of high toxicity of the substances in cell cultures. Owing to their structure or stability, a variety of biotechnological products cannot be subjected to expensive purification or inactivation using product-damaging or cytotoxic antiviral substances such as solvents.
It was therefore the object of the invention to provide a mild method of inactivating lipid-enveloped viruses in biological or biotechnological products and cell cultures, which method enables to render these products or cell cultures free of viruses in an exceedingly rapid and effective fashion, with no denaturing of products or impairing the cell cultures in their productivity. Also, said method should allow the treatment of heat-labile products and avoid substances having in vivo toxicity.
The object of the invention is accomplished by using cyclic peptides containing &bgr;-hydroxyfatty acids and &bgr;-aminofatty acids (lipopeptides). These lipopeptides were found to have a surprisingly high inactivation potential for lipid-enveloped viruses, and thus, they are excellent for use in accomplishing the object of the invention. Part of these lipopeptides were found to be substantially more effective compared to synthetic surfactants used in virus inactivation up to now. In addition, they readily undergo biological degradation and have a substantially lower in vivo toxicity than synthetic surfactants. Compared to conventional antiviral substances, the lipopeptides used according to the invention have the advantage of thermal stability and good water-solubility.
From the literature, two [Ile
7
] and [Leu
7
] surfactins which belong to the lipopeptides, are known to exhibit moderate anti-HIV-1 activity (H. Itokawa et al., Chem. Pharm. Bull. 42, 604-607 (1994)). In Journal of Antibiotics, Japan XLIII 267-280 (1989), N. Naruse et al. describe pumilacidins as antivirally effective against herpes simplex virus (HSV-1). Neither of these papers indicates the considerable inactivation potential of these substances, allowing extensive inactivation of lipid-enveloped viruses at low concentrations within an exceedingly short time.
Thus, the inactivation method according to the invention is characterized in that the biological or biotechnological products are added with a lipopeptide or a salt or ester thereof, or a mixture of lipopeptides or salts or esters thereof at an overall concentration of 1-100 &mgr;M, preferably 1-80 &mgr;M, and that the inactivation is performed at room temperature within from 30 min to 2 hours at maximum, with about 99% of the viruses already being inactivated after 30 min. As a result of the exceedin
Kamp Roza Maria
Pauli Georg
Vater Joachim
Vollenbroich Dirk
Birch Stewart Kolasch & Birch, LLP.
Housel James
Lucas Zachariah
LandOfFree
Inactivating process for lipid envelopped virus, and new... does not yet have a rating. At this time, there are no reviews or comments for this patent.
If you have personal experience with Inactivating process for lipid envelopped virus, and new..., we encourage you to share that experience with our LandOfFree.com community. Your opinion is very important and Inactivating process for lipid envelopped virus, and new... will most certainly appreciate the feedback.
Profile ID: LFUS-PAI-O-3195191