Claims
- 1. Interleukin-2 containing liposomes.
- 2. Liposomes according to claim 1 wherein the interleukin is human IL-2.
- 3. Liposomes according to claim 1 wherein the interleukin is IL-2-serine 125.
- 4. Liposomes according to claim 1 wherein the interleukin is IL-2 A-1, IL-2 A-2, IL-2 B-1 or IL-2 B-2.
- 5. Liposomes according to claim 1 wherein the interleukin is IL-2 Gln-26, IL-2 Phe-121 or IL-2 Stop-121.
- 6. The liposomes of claim 1 in which the lipid content consists essentially of phospholipids.
- 7. The liposomes of claim 1 in which the lipid content consists essentially of phospholipids and steroid lipids in a weight ratio of phospholipids to steroid lipids of from 6:1 to 1:1.
- 8. The liposomes of claim 1 in which the interleukin-2 is present in an amount of 20 to 200 micrograms per milliliter of the aqueous phase and the lipid concentration is present in an amount of 10 to 100 milligrams per milliliter of the aqueous phase.
- 9. Lyophilized interleuken-2 containing liposomes.
- 10. The liposomes of claim 2 in which the lipid content consists essentially of phospholipids.
- 11. The liposomes of claim 2 in which the lipid content consists essentially of phospholipids and steroid lipids in a weight ratio of phospholipids to steroid lipids of from 6:1 to 1:1.
- 12. The liposomes of claim 2 in which the interleukin-2 is present in an amount of 20 to 200 micrograms per milliliter of the aqueous phase and the lipid concentration is present in an amount of 10 to 100 milligrams per milliliter of the aqueous phase.
- 13. Lyophilized human interleuken-2 containing liposomes.
- 14. The method of stimulating the T-cell dependent immune response in a mammal in need of such treatment comprising intravenously administering to such mammal a T-cell dependent immune response stimulating effective amount of interleukin-2-containing liposomes.
- 15. The method of claim 14 in which the interleukin-2-containing liposomes are human interleukin-2-containing liposomes.
Priority Claims (7)
| Number |
Date |
Country |
Kind |
| 8409125 |
Apr 1984 |
GBX |
|
| 8409126 |
Apr 1984 |
GBX |
|
| 8409127 |
Apr 1984 |
GBX |
|
| 8409128 |
Apr 1984 |
GBX |
|
| 8420381 |
Aug 1984 |
GBX |
|
| 8423700 |
Sep 1984 |
GBX |
|
| 8423701 |
Sep 1984 |
GBX |
|
Parent Case Info
This is a continuation of application Ser. No. 896,551, filed Aug. 14, 1986, now abandoned, which in turn is a continuation of application Ser. No. 721,316, filed Apr. 9, 1985, now abandoned.
This invention relates to interleukin therapy. Interleukin-2 (hereinafter IL-2) is a naturally occurring protein factor first discovered in 1976. It is produced from T-cells activated by an antigen or lectin and is a factor essential for the proliferation of T-cells. IL-2's of various structures have been isolated from a number of animal species, e.g. mouse, primate such as the gibbon, ape, and human. Human and other IL-2's have been purified from various sources such as peripheral blood lymphocytes, tonsilar lymphocytes, spleen lymphocytes, T-cell leukemia and T-cell hybridoma cultures.
As indicated above IL-2 is a factor essential for the proliferation of T-cells, themselves intimately involved in the body's immune response mechanisms. IL-2 is anticipated to have therapeutic potential of immense proportions. In particular, it may have value in the therapy of tumours since it leads to proliferation of antigen specific T-cells e.g. against tumour antigens and these T-cells can inhibit tumour growth. It is also known that IL-2 induces production of gamma-interferon and activates natural killer cells. It is also expected that IL-2 will have a variety of applications against immunological disorders, such as neoplastic diseases, bacterial or viral infections, immune deficient diseases, auto-immune diseases etc (see B. Papermaster et al, Adv. Immunopharm. (1980) 507).
As with other naturally occurring products such as the interferons, before the advent of genetic engineering, IL-2 was available in small quantities only.
However, following methodology similar to that previously published for other proteins (e.g. gamma-interferon) the structure determination and expression of a cloned gene for human IL-2 was effected (see S. Taniguchi et al, Nature (1983) 302, p. 305-310 and European Patent Publication No. 91539 the contents of both which are incorporated by reference. This particular IL-2 is referred to hereinafter as known IL-2.
On the basis of the nucleotide coding sequence the amino acid sequence of the human IL-2 polypeptide, namely that comprising the amino acids numbered 1 to 153 in FIG. 3a of the above Nature article, was deduced. It was postulated, however, that the first 20 animo acids of this sequence could be cleaved on transmembrane transport. The mature human IL-2 would then consist of 133 amino acids beginning with the Ala at position 21.
This structure of human IL-2 has since been confirmed in our and other laboratories.
It is also possible by using recombinant DNA techniques to produce IL-2's of different structures. For example, modifications of the human IL-2 polypeptide having one or more amino acids absent or replaced by other amino acids, e.g. as described in the above mentioned European Patent Publication, in particular pages 23 to 25 thereof, may be produced by correspondingly modifying the human IL-2 gene. For example, if desired Cysteine residues may be replaced by other amino acids, e.g. serine, as described in Cetus European Patent Publication No. 109748 and Belgium Patent 898016, e.g. IL-2-serine-125, the contents of which are hereby incorporated by reference.
Further examples of such interleukins are disclosed in PCT patent publication WO 85/00817, the contents of which are incorporated herein by reference. These include IL-2 Gln-26, IL-2 Phe-121, and IL-2 Stop 121.
Furthermore, IL-2's from different sources such as the gibbon ape may be produced by recombinant DNA techniques and modification of these may also be produced in similar manner. Furthermore, IL-2's may, as indicated, be isolated (albeit in relatively small quantities) from natural sources and may differ from the products produced by recombinant DNA technology in for example glycosylation. Moreover, allele variantes may be produced.
IL-2's may also be produced by cultivating human cells, e.g. in the presence of an inducer. Examples are IL-2 A-1, IL-2 A-2, IL-2 B-1, IL-2 B-2 as defined in Danish Patent Application No. 3317/84 and European Patent Publication No. 132359, the contents of which are incorporated herein by reference.
By "interleukin" as used hereinafter is meant any polypeptide, including but not limited to those described above, whether isolated from natural sources or produced by a synthetic or biosynthetic method and which has IL-2 activity. In accordance with the present invention, the preferred interleukin is human IL-2 or a modification thereof, preferably as produced by recombinant DNA methods. Trials so far carried out with interleukin have been by injection, e.g. intravenously. These have shown for example that human IL-2 has a very short half-life of under 5 minutes. There is clearly a need for a sustained release parenteral form of interleukin providing a sufficiently long duration of action, however, up to now little has been published on specific galenic formulations on interleukins.
Furthermore, trials carried out in our laboratories have shown that interleukin may be incorporated into liposomal forms, which on administration, e.g. intravenous administration, still show IL-2 activity. Furthermore, the liposomes of the present invention have a prolonged half-life, are passively targetted into the spleen, lung, bone marrow or lymph nodes.
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| Number |
Date |
Country |
| 109861 |
May 1984 |
EPX |
| 132359 |
Jan 1985 |
EPX |
Non-Patent Literature Citations (1)
| Entry |
| Ostro. "Liposomes", Marcel Dekker, Inc., pp. 73, 242, 243 and 315. |
Continuations (2)
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Number |
Date |
Country |
| Parent |
896551 |
Aug 1986 |
|
| Parent |
721316 |
Apr 1985 |
|
Patent Grant
4863740
