Drug – bio-affecting and body treating compositions – Inorganic active ingredient containing – Heavy metal or compound thereof
Reexamination Certificate
2001-08-20
2004-03-23
Russel, Jeffrey E. (Department: 1654)
Drug, bio-affecting and body treating compositions
Inorganic active ingredient containing
Heavy metal or compound thereof
C514S034000, C514S045000, C514S049000, C514S249000, C514S283000, C514S564000, C536S006400, C536S016800, C536S017200, C536S027210, C536S028400, C544S261000, C546S051000, C562S449000
Reexamination Certificate
active
06709679
ABSTRACT:
FIELD OF THE INVENTION
This invention relates to tumor-inhibiting conjugates of proteins and polymers consisting of a suitable carrier system and cytostatic compounds. Further, the invention relates to methods for the production of such conjugates and the use of these. Immuno-conjugates or conjugates of protein or of polymer are compounds which consist of a suitable carrier substance, such as, for example, an antibody, a growth factor, a structure similar to hormones or peptides, a protein or a polymer, and one or more cytotoxic active substances such as, for example, cytostatics, toxins or radioactive isotopes. The carrier substances have, as a rule, the characteristic of preferably accumulating in the tumor tissue, so that in this way also the active substance bound to the carrier substance accumulates in the tumor tissue and thus a selective anti-tumor therapy is achieved. Chemoimmuno-conjugates are conjugates of carrier substances and cytostatic compounds, wherein the carrier, as a rule, is an antibody.
PRIOR ART
The cytostatics currently used against cancers have a series of strong systemic side-effects and do not exhibit accumulation in the tumor tissue, so that new derivatives and formulations are being researched which make selective anti-tumor therapy possible. For this purpose. chemoimmuno-conjugates or conjugates of proteins or of polymers consisting of one suitable carrier substance or cytostatics are being developed.
As carrier substances, among others, antibodies, growth factors, serum proteins, structures similar to hormones or peptides, or polymers are considered, for which, as a rule, an accumulation in the tumor tissue is known (Mägerstädt, M.: Antibody Conjugates and Malignant Disease, Library of Congress 1990: Chadwick, C. M.: Receptors in Tumour Biology, Cambridge University Press, 1984, Seymour, L. W.
CRC Crit. Rev. Ther. Drug Carrier Sys.
(1992), 9, 135-187; Maeda, H.; Matsumura, Y.
CRC Crit. Rev. Ther. Drug Carrier Sys.
(1989), 6, 193-210). The present invention comprises human serum transferrin and serum albumin as carrier proteins, of which the accumulation in the tumor tissue is documented (Ward, S. G. Taylor, R. C.: 1-54, in Metal-Based Drugs (Gielen, M. F. (Ed.)), Freund Publishing House Ltd, 1988; Sinn, H., Schrenk, H. H., Friedrich, A., Schilling, U. and Maier-Borst, W. (1990),
Nucl. Med Biol.
Vol. 17(8), 819-827) as well as polyethylene glycols (PEGs) as carriers of cytostatic compounds (Topchieva, I. N. (1990), Polym. Sci. USSR 32, 833-851; Poly(ethylene glycol) Chemistry: Biotechnical and Biomedical Applications (1992), Ed. J. M. Harris, Plenum Press, New York). PEGs are, due to their bio-compatibility, their good water-solubility and synthetic divergence, very suitable for the development of therapeutic polymer conjugates. In recent years, PEGs have been conjugated mainly with medically significant proteins and enzymes (Overview in Topchieva, I. N. (1990), Polym. Sci. USSR, 32, 833-851). The production of chemoimmuno-conjugates and conjugates of proteins or of polymers occurs generally either through direct coupling of carrier substance and active substance or with the help of spacer groups, so-called homo- or heterobifunctional reagents. Until now, mainly the method of direct coupling has been used which, however, often leads to polymeric products and not-unequivocally-defined conjugates. Recently, several chemoimmuno-conjugates (European Patent Application EP
91-117535 911615,
European Patent Application EP
90-109268 900516,
PCT International Patent Application WO 90-CA251 900809, British (UK) Patent Application GB 83-5104 830224 and European Patent Application EP
89-102370 890210
), which were produced using specific bifunctional reagents, were suggested as cytostatically effective media. Furthermore, from DE 41 22 210 A1, conjugates of tumor-active compounds with transferrin or albumin are known, wherein the tumor-active compound is activated with N-hydroxy succinimide and carbodiimide and the thus-obtained mixture is directly coupled to the carrier protein.
DESCRIPTION OF THE INVENTION
It has now been found that conjugates of transferrin, albumin and polyethylene glycol, consisting of transferrin, albumin and polyethylene glycol, with a mass of between 5000 and 200000 Da and, at least, one cytostatic compound derivatized through compounds of maleinimide or N-hydroxysuccinimide, have a tumor-inhibiting effectiveness which is equal or higher than that of the cytostatic compound. Suitable for the production of these conjugates of protein or polymer are cytostatic compounds such as the anthracyclines, doxorubicin, daunorubicin, epirubicin, idarubicin and mitoxandrone, the alkylates, chloroambucil and melphalan, the antimetabolites, methotrexate, 5-fluorouracyl, 5′-desoxy-5-fluorouridine and thioguanine, the taxoides, paclitaxel and docetaxel, the camptothecins, topotecan and 9-aminocamptothecin, the podophyllotoxin derivatives, etoposide, teniposide and mitopodoside, the vinca alkaloids, vinblastine, vincristine, vindesine and vinorelbine and a compound of the general I, II, III or IV:
n=0-6, X=—NH
2
, —OH, —COOH, —O—CO—R—COR*, —NH—CO—R—COR*, wherein R is an aliphatic carbon chain with 1-6 carbon atoms or a substituted or unsubstituted phenylene group and R* H, phenyl, alkyl with 1-6 carbon, and the amine functions are provided with a protective group such as the tert.-butyloxycarbonyl protective group, which were derivatized with a compound of maleinimide or N-hydroxysuccinimide. In doing so, the cytostatic compounds are, as a rule, reacted with a maleinimide compound or N-hydroxysuccinimide compound which has at least one functional group which is suitable for binding to the cytostatic, such as an amino, hydroxy, carbonic acid, carbonic acid chloride, sulfonic acid, sulfonic acid chloride, acid hydracide, or hydrazino, oxycarbonyl chloride, aldehyde or keto group, so that maleinimide derivatives or N-hydroxysuccinimide ester derivatives of cytostatic compounds are prepared, wherein the chemical linkage between the maleinimide compound and cytostatic compound occurs through an amide, ester, imine, hydrazone, carboxyl hydrazone, oxycarbonyl, acetal or ketal bond. In the maleinimide or N-hydroxysuccinimide compounds which are obtained from the compounds of the formulas I-IV, the cytostatic cis-configured platinum unit is introduced subsequently, that is, the corresponding platinum(II)-complexes are obtained, after removal of the protective group, either through reaction with a tetrachloroplatinate salt or with cis-[PtA
2
B] (A=halogen, B=(NH
3
)
2
, ethylene diamine, propane diamine, 1,2-diaminocyclohexane).
Through reacting the derivatized cytostatic compounds with native or thiolated transferrin or albumin or with hetero- or homobifunctional PEGs with a mass of between 5000 and 200 000 Da—Overview 1:
Overview 1:
Heterobifunctional PEGs
Hommobifunctional PEGs
conjugates of proteins or polymers are prepared which are produced simply and effectively, having a high purity, have an excellent water-solubility in comparison to several of the original cytostatic compounds, are stable formulations in a physiologic buffer and which have an in vitro anti-proliferation effectiveness against human tumor cells which is equal to or better than that of the unbound cytostatics. Furthermore, the conjugates exhibit a equal good or improved anti-tumor effectiveness in vivo and an improved tolerability. The conjugates of protein or polyethylene glycol realized through such couplings, which are very suitable for a selective treatment of cancer diseases, are object of the invention and are described in the following.
The method for the synthesis of conjugates of protein or polyethylene glycol occurs in the conjugates with maleinimide derivatives in four steps (Steps 1 to 4), in the conjugates with N-hydrosuccinimide ester derivatives in three steps (Steps 1, 2 and 4):
Step 1: Synthesis of maleinimide or N-hydroxysuccinimide compounds
Step 2: Synthesis of maleinimide derivatives or N-hydroxysuc
Burns Doane Swecker & Mathis L.L.P.
Russel Jeffrey E.
LandOfFree
Antineoplastic conjugates of transferin, albumin and... does not yet have a rating. At this time, there are no reviews or comments for this patent.
If you have personal experience with Antineoplastic conjugates of transferin, albumin and..., we encourage you to share that experience with our LandOfFree.com community. Your opinion is very important and Antineoplastic conjugates of transferin, albumin and... will most certainly appreciate the feedback.
Profile ID: LFUS-PAI-O-3202042