US20030017987A1

US20030017987A1 - Therapeutic, prophylactic, and diagnostic agent for cancer, useful for characterizing cancer cells with individual properties

Info

Publication number: US20030017987A1
Application number: US10/238,726
Authority: US
Inventors: Michael Zeppezauer; Hans-Peter Leinenbach; Reiner Class; Cordula Fassbender
Original assignee: Individual
Current assignee: Individual
Priority date: 1997-04-11
Filing date: 2002-09-11
Publication date: 2003-01-23
Also published as: US7902146B2; US20070203067A1

Abstract

A therapeutic or prophylactic agent for cancer is disclosed which damages the membrane and kills cancer cells, in particular of the blood-forming system, having membrane protein aggregates which contain several core histones or largely core-like histones and/or their parts. The therapeutic or prophylactic agent contains at least one pure histone or its active sequence section selected from the group composed of histone H1, H1 subtypes, H2A, H2B, H2A:H2B dimer, H3 and H4, covalent modified histones of the above-mentioned type and/or their active sections and functionally and structurally similar proteins (protamines, histone-like proteins of prokaryotic and archae bacteria).

Description

The invention relates to histones and/or histone-like proteins (i.e. protamines, bacterial histone-like proteins) as a therapeutic and prophylactic agent against cancer cells and is a continuation of application Ser. No. 09/402,468, the entire disclosure of which is herein incorporated by reference.[0001]

According to Reiner Class et al. (in American Journal of Clinical Oncology (CCT) 19 (5) 1996, pp. 552-531), the effect of histones on cancer cells is due to the fact that those cancer cells, which are killed by histone H1, express a so called receptor protein in their membranes that is able to bind histone H1. Said receptor protein has been isolated by affinity chromatography using immobilized histone H1. It has been determined by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) that the receptor protein has an electrophoretic mobility corresponding to a molecular mass of approximately 33,000 Da (33 k Da) ±2,000 Da.

The binding of histone H1 to the membrane-embedded receptor protein causes a destruction of the membrane integrity, resulting in an uncontrolled efflux of soluble cellular components such as proteins, ions and other molecules and influx of other substances, ultimately resulting in cell death.

The invention is based on the rationale that tumor cells of different histological origin (e.g. leukemia, lymphomas, sarcomas, carcinomas, melanomas etc.) possess special individual properties and that a better understanding of said individual and cell line-specific properties can offer a starting point for a novel and effective therapy. It could be demonstrated that the receptor protein is not characteristic for a single or several distinct cell types but rather represents an individual, cell line-characteristic property that can be made accessible for a targeted therapy.

The invention has set its ultimate goal to obtain more knowledge about the receptor protein, translating into a novel teaching for therapeutic and prophylactic treatments or suppressions of individual cancer cells expressing such receptor proteins.

Therefore, the invention also comprises a diagnostic tool for the recognition of those cancer cells that express (as an individual characteristic) such receptor proteins, which makes the success of the teaching for the therapeutic treatment of said individual cancer cells (i.e. the outcome) predictable. This implies that cancer cells can be classified in a novel way depending on the success of the treatment with the therapeutic agent (i.e. histone H1). As soon as reliable data become available that can predict the predisposition of distinct people to develop cancer cells with said individual properties, the therapeutic (that is subject of this invention) can also be used in a prophylactic regimen. This will be especially applicable if the natural level of histones in the blood of these people has been recognized as too low.

This invention is founded on the observation that above receptor protein in the membrane of individual cancer cells (especially cancer cells of the hematopoietic system such as leukemias, lymphomas and myelomas) contains several histone proteins or histone-like polypeptides and/or parts thereof or is largely composed of them. These proteins can bind and cross link with externally added histones or histone-like proteins (e.g. protamine and bacterial histone-like proteins), whereby the regimen can be of therapeutic or diagnostic nature. Appropriate antibodies that recognize the receptor proteins can also be utilized as diagnostic tools.

The invention is characterized by the content of

claims

1, 24, 25, and 26. Advantageous executions result from the characteristics of the claims and from the following description.

In the description, the figures show respectively: [0009]
FIG. 1 Recombinantly produced fragments of histone H5; [0010]
FIG. 2 examples 1 to 5, regarding the cytotoxicity of histone H1 and recombinant histone fragments towards acute myeloid leukemia; [0011]
FIG. 3 examples 6 to 10, regarding the cytotoxicity of histone H1 and recombinant histone fragments towards cell line IM9; [0012]
FIG. 4 examples 11 to 15, regarding the cytotoxicity of histone H1 and recombinant histone fragments towards cell line OH77; [0013]
FIG. 5 examples 16 to 18, regarding the cytotoxicity of histone H1 recombinant histone fragments towards cell line Daudi; [0014]
FIG. 6 photographs A and B of untreated cells of cell line OH77 at different magnifications; and [0015]
FIG. 7 photographs C and D tumor cells treated with histone H1.[0016]

Histone-like proteins are defined here as protamines and distinct bacterial proteins. Some of the histone-like proteins of bacterial origin are listed in table 1 including relevant references from the literature. The similarity between the listed bacterial histone-like proteins with histones has been determined from a structural point of view by X-ray crystallography on protein crystals, from a functional point of view by DNA-binding and transcription studies, and from a biochemical point of view by sequence homologies with histone H1.

TABLE 1


Protein	BaCteria	Reference

HMf	Methanothermus	Ronimus, R. S. and Musgrave, D. R. (1996)
HAN1	fervidus	Purification and characterization of a histon-
HAN2		like protein from Archaeal isolate AN1,
		a member of the Thermococcales. Mol.
		Microbiol., 20:77-86
HAN1	Thermococcus	Ronimus, R. S. and Musgrave, D. R. (1996)
	spec.	A gene, han1A, encoding an archaeal
		histone-like protein from the Thermococcus
		species AN1:homology with eukaryal
		histone consensus sequences and the
		implications for delineation of the
		histone fold. Biochim. Biophys.
		Acta, 1307:1-7
HMfA	Methanothermus	Decanniere, K., Sandman, K., Reeve,
HMfB	fervidus	J. N., Heinemann, U. (1996)
		Crystallization and preliminary X-ray
		characterization of the Methanothermus
		fervidus histones MMfA and HMfB.
		Proteins, 24:268-71
Hc1	Chalmydia	Petersen, L. B., Birkelund, S., and
Hc2	trachomatis	Christiansen, G. (1996)
		Purification of recombinant Chalmydia
		trachomatis histone H1-like protein Hc2,
		and comparative functional analysis of Hc2
		and Hc1.
		Mol. Microbiol., 20:295-311
MC1	Methanosarcina	Teyssier, C., Toulme, F., Touzel, J. P.,
	spec.	Gervais, A., Maurizot, J. C., and Culard, F.
		(1996)
		Preferential binding of the archaebacterial
		histone-like MC1 protein to negatively
		supercoiled DNA minicircles.
		Biochemistry. 35:7954-7958
BpH1	Bordetella	Scarlato, V., Arico, B., Goyard, S., Ricci,
	pertussis	S., Manetti, R., Prugnola, A., Manetti, R.,
		Polverino-De-Laureto. P., Ullmann, A.,
		and Rappuoli, R. (1995)
		Mol. Microbiol., 15:871-881

Initially, the above mentioned receptor protein in the membrane of individual cancer cells has been characterized by the inventors. [0018]
The mass spectrometric analysis of the protein bands derived from SDS-PAGE using MALDI-MS technique (matrix-assisted laser desorption/ionization mass spectrometry) revealed that the electrophoretically homogeneous band contained three different proteins with the molecular masses of 11.2, 13.7, and 15.0 kDa. [0019]
In detail, the following mean values have been obtained:[0020]
11,175±40 kDa
13,730±40 kDa and
15,035±80 kDa
The mean values are calculated from single- and double charged molecule ion signals and from 4 to 6 sum-spectra, acquired from the examined samples. [0021]
In comparison to above molecular masses of the receptor polypeptides, the below listed human histone sequences have the following molecular masses:[0022]
H4 11,282 kDa
H2B 13,774 kDa
H3 15,324 kDa and
H2 14,004 kDa
The sequence analysis by automated Edman protein degradation identified a peptide sequence that corresponded to [0023] amino acid residues 23 to 59 of human histone H4, differing only in position 23 and 35. This represents a 97.2% homology compared to the known sequence of histone H4 in healthy cells.
These findings justify the surprising conclusion that the mentioned receptor protein consists of or contains those proteins that can be related to the histones H4, H2B, and H3 or to proteins very similar to them. The presence of histone proteins in the membrane of individual cancer cells is most likely due to the tumorigenic transformation which these cells undergo, likewise their quantitative composition and potential structural differences compared to the nuclear histones. [0024]
According to the invention, the histone proteins belonging to the histones H2A, H2B, H3 and/or H4 (the so called core histones) that are expressed in the cell membrane of individual cancer cells at comparatively high concentrations can be cross-linked by the external addition of histones, especially histone H1 or of active components thereof. The cross-linking causes a loss of membrane integrity in the particular cancer cell, ultimately resulting in cell death. Mentioned histone proteins can be of natural origin, e.g. extracted from bovine tissue, or can be recombinant proteins. Covalently modified histone proteins or parts thereof or functionally and structurally similar proteins can also be used. Suitable covalent modifications include but are not restricted to derivatization with polyoxyethylene chains (“pegylation”). An example for functionally and structurally similar proteins is protamines or histone-like proteins from prokaryotic organisms or archebacteria. [0025]
It is a known that histone H1 molecules serve as linker between the so-called nucleosomes, a well-organized complex of core histones, thus forming the condensed structure of the chromatin. To achieve this, largely intact histone H1 molecules are required, containing a compact and folded central part (i.e. the globular domain) with the adjacent flexible N- and C-terminal domains. [0026]
One could deduct from this knowledge a method to pinpoint the biologically active (i.e. cytotoxic) domains of the histone H1 molecule that still guarantees the cross-linking with or binding to those core histones that are expressed in cell membranes of individual cancer cells. [0027]
For this, recombinantly manufactured histone H5 has been used partly in cytotoxicity experiments. Histone H5 is derived from chicken erythrocytes and represents the erythrocyte-specific variant of histone H1. Fragments of this histone protein have been employed partly in which either both terminal tails have been extensively shortened or in which the C-terminal tail has been gradually shortened. These fragments and their manufacturing by genetechnology methods have been described by Gerchman et al. in Protein Expression & Purification, 5(1994), pages 242-251. [0028]
FIG. 1 shows the employed H5-fragments and their structural composition. They are: the whole histone H5 molecule (H5), essentially the globular domain only (GH5), the globular domain with the N-terminal domain (NGH5), the globular domain with the N-terminal domain and the C-terminal amino acids 98-131 (NGC1H5), and finally, the globular domain with the N-terminus and the C-terminal amino acids 98-151 (NGC2H5). All proteins and culture media used have been tested negative for endotoxin. [0029]
FIG. 1 as a graphic representation of the three structural domains of the histone molecules H5 (GH5, NGH5, NGC1H5, and NGC2H5) describes recombinantly produced histone fragments. [0030]
To investigate potential structure-function relationships of the effect of calf thymus-derived histone H1 on transformed cancer cells, a series of experiments has been performed, comparing histone H1 with above listed recombinant polypeptide fragments of histone H5. The following cells/cell lines have been tested: a primary cell line derived from the peripheral blood of a female patient with acute myeloid leukemia (AML), cells of the leukemia cell line IM9, and cells of the lymphoma cell line OH77. In all experiments, histone H1 was cytotoxic at a concentration of 250 ig/ml. The cytotoxic effects of the longer recombinant H5-derived polypeptides, NGC1H5 and NGC2H5, towards cells of the AML and the lymphoma cell line were comparable to histone H1 whereas stronger cytotoxic effects could be observed for the cell line IM9. The smallest growth inhibitory effects have been induced in all cell types by the globular domain GH5 and the fragment NGH5. The recombinant fragments have been used in concentrations equimolar to 250ig/ml histone H1. The results are summarized in the following examples 1-5. [0031]
FIG. 2 shows the cytotoxicity of histone H1 and the recombinant histone fragments towards leukemia cells of a female patient with AML (acute myeloid leukemia). [0032]
In examples 1 to 5, 1×10[0033] ⁶cells/ml have been incubated for 48 h under the following standard conditions:
H1: Histone H1; recombinant fragments of histone H5: GH5: fragment GH5; NGH5: fragment NGH5; C1: fragment NGC1H5; C2: fragment NGC2H5. Histone H1 and the recombinant fragments have been employed at 12 iM. The viability of the cells has been determined using the MTT method. Shown are the mean values of 4 parallel experiments. [0034]
In FIG. 3 and in examples 6 to 10, the cytotoxic effects of histone H1 and recombinant histone fragments are shown for cells of the myeloid leukemia cell line IM9. [0035]
Furthermore, in FIG. 4 and examples 11 to 15 show the cytotoxic effects of histone H1 and recombinant histone fragments for cells of the lymphoma cell line OH77. [0036]
In examples 6 to 15, 5×10[0037] ⁴cells/ml have been incubated for 48 h under the following standard conditions:
H1: Histone H1; recombinant fragments of histone H5: GH5: fragment GH5; NGH5: fragment NGH5; C1: fragment NGC1H5; C2: fragment NGC2H5. Histone H1 and all recombinant H5 fragments have been incubated at 12 iM. The viability of the cells has been determined using the MTT method. Shown are the mean values of 4 parallel experiments. [0038]
From the examples it follows that the globular domain of the histone H1 (or H5) molecule GH5 possesses only a small or no cytotoxic effects at all (indicated by a cytotoxicity) index smaller than 20). Similar is valid for the fragment NGH5. which almost completely lacks the C-terminal domain. If at least the [0039] residues 108 to 131 are present and if the N-terminal domain is intact (NGC1H5), a fragment results whose cytotoxic activity reaches or even surpasses that of intact histone molecule (see examples 4 and 9). The same is true for NGC2H5. which is lacking only 38 amino acids of the C-terminal domain.
Similar relationships count respectively for histone fragments with shortened N-terminal domain and intact C-terminal domain. [0040]
According to the invention, it suggests itself to use human histone H1 subtypes instead of histone H5, especially the subtypes H1.1, H1.2, H1.3, H1.4, and H1.0 or parts thereof. [0041]
FIG. 5 and examples 16 and 17 show the cytotoxic activity of recombinantly produced human (rh) histone H1 subtypes H1.0 and H1.2 in comparison to calf thymus-derived (bovine) histone H1 towards cells of the burkitt lymphoma cell line Daudi (FIG. 5, example 18). [0042]

The following table 2 lists the mentioned human histone H1 subtypes with their respective N- and C-terminal domains. The corresponding globular domain between the N- and C-terminal domains has not been listed here. In analogy to the results obtained with the histone H5 from chicken, it is reasonable to expect that these subtypes exert similar cytotoxic effects towards those cancer cells that contain membrane-standing core histone receptors.

TABLE 2


N-terminal	C-terminal	Necessary N-	Necessary C
domain	domain	terminal domain	terminal domain

H1.1 1-40	118-214	from 16; 20-26	118-138
H1.2 1-38	105-212	from 16; 20-26	105-125
H1.3 1-39	117-220	from 16: 20-26	117-137
H1.4 1-38	116-218	from 16; 20-26	116-136
H1.0 1-25	99-193	from 11; 16-20	99-119

In this above listed gradation, the C- and N-terminal domains have been defined as those areas that contain the amino acid proline (P) in their sequences, because proline prohibits the formation of helices by causing a stretching of the polypeptide chain. [0044]
In the literature, the borders of the domains are defined differently. In the case of the chicken histone H5, some authors define the N-terminus as [0045] amino acids 1 to 18, and the globular domain as amino acid 19 to 108, whereas Gerchman et al. (in the preceding reference) define the globular domain as amino acids 24 to 96.
In summary, one can conclude that the proteins of the individual receptor of cancer cells—based on current knowledge—represent an aggregate of the core-histones H2A, H2B, H3 and/or H4 or that it contains such histones or core histone-like proteins. Whether DNA is present on the membrane surface cannot be determined conclusively. However, it cannot be excluded. [0046]
Additionally, it cannot be excluded that protamines can be synthesized in cancer cells as a consequence of their neoplastic transformation. These protamines can then be localized in the cell membrane instead of the core histones or together with the core histones, forming complexes with and binding to histones or histone-like proteins. [0047]
Histones belonging to the nucleosome (i.e. core histones) are connected to each other tightly by non-covalent forces thus mimicking a higher molecular weight. They are also associated with each other in the cell membrane where they end up due to a transformation-dependent disturbance in the regulation. [0048]
The mechanism of action of the therapeutic agent, which is subject of this invention, is based-on the cross linking of membrane-standing histone proteins through (a) externally administered histone proteins, especially histone H1 and parts thereof that can be cross linked, (b) biologically active, covalently modified histones or (c) through functionally and/or structurally histone-like proteins. As a consequence of this cross-linking or binding, larger aggregates are formed within the cell membrane. These aggregates have a pore-like character, give raise to the formation of pores or cause mechanical deformation of cell membrane components that are directly associated with structural components of the cytoskeleton. These events result in the disruption of the cell membrane integrity and, ultimately, cell death. It is possible that signal transduction mechanisms and apoptotic processes are involved in this but they are not the determining process that leads to cell death. [0049]
The biological activity of the therapeutic agent, which is subject of this invention, is made clear in photographs A and B of FIG. 6 and photographs C and D in FIG. 7. [0050]
In FIG. 6, photograph A shows untreated cells of the lymphoma cell line OH77 (magnification 2,000-fold), photograph B is a detail from photograph A at a 20,000-fold magnification. Through this, the typical surface structure of an intact lymphatic tumor cell is clarified. [0051]
In FIG. 7, photograph D depicts the drastic change of the tumor cells after 12 hours incubation with 200 ig/ml histone H1. It becomes evident that the cells contract to a sphere-like shape and, after a 24 hours incubation time with histone H1, completely disintegrate (as shown in photograph C of FIG. 7). [0052]
The pure histone H1 preparation has been employed at the above listed concentration and was resuspended in normal media used for cell culture as described in the previously listed reference. [0053]
The invention also includes a method for the diagnosis of individual cancer cells expressing the described receptor protein in their membrane. According to the invention, those histones—especially histone H1 or active parts thereof or suitable antibodies—will be used for diagnosis that bind to the receptor protein on cancer cells thus allowing for the first time a classification of individual cancer cells containing this particular receptor protein. The diagnostic techniques needed for this are widely known and state-of-the-art. The same is valid for a technique to find suitable antibodies, which recognize the receptor protein itself in the membrane of cancer cells or related structures formed by aggregation. [0054]

Claims

We claim:

1. A cancer-therapeutic agent on the basis of histones or histone-like proteins (e.g. protamines as well as histone-like proteins from prokaryotes and/or archebacteria), especially histone H1 or active parts thereof, for the treatment of cancer cells, especially those of the hematopoietic system, having individual membrane-embedded receptor proteins that either contain different histones or histone-like polypeptides and/or parts thereof or are largely composed of them.

2. A therapeutic agent according to claim 1, wherein the agent is characterized by its reaction with a receptor protein in the membrane of cancer cells. Said receptor protein contains or consists of at least three different polypeptides, which belong to individual core histones or parts thereof.

3. A therapeutic agent according to claim 2, wherein the agent is characterized by three different molecular masses in the range of 11 kDa, 14 kDa, and 15 kDa, especially 11,715±40 Da, 17,730±40 Da, and 15,035±80 Da.

4. A therapeutic agent according to one of the previous claims, wherein the agent is characterized by a receptor protein containing a peptide sequence that has homology greater than 90% to the sequence 23-59 of human histone H4 (molecular mass 11,282 Da).

5. A therapeutic agent according to claim 4, wherein the agent is characterized by a receptor protein containing two peptide sequences (additionally to the histone H4 sequences) with high homology to parts of human histone H2B (molecular mass 13,774 Da) and human histone H3 (molecular mass 15,324 Da).

6. A therapeutic agent according to claim 1 or 2, characterized by a receptor protein that contains or is composed of core histones or core histone-like polypeptides in aggregates similar to or alike nucleosomes.

7. A therapeutic agent according to claim 1, wherein the agent is characterized by containing at least one histone protein or biologically active sequence parts thereof, selected out of a group of proteins consisting of one histone H1, the pure histone H1 subtypes, H2A, H2B, H2A:H2B dimer, H3, and H4.

8. A therapeutic agent according to claim 7, wherein the agent is characterized by containing histone H1 or a biologically active part thereof.

9. A therapeutic agent according to claim 8, wherein the agent is characterized by the fact that the biologically active histone H1 part is defined as the globular histone H1 domain in connection with the complete N-terminal domain or a sequence part thereof.

10. A therapeutic agent according to claim 8, wherein the agent is characterized by the fact that the biologically active histone H1 part is defined as the globular histone H1-domain in connection with the C-terminal domain or a sequence part thereof.

11. A therapeutic agent according to claim 8, wherein the agent is characterized by the fact that the biologically active histone H1 part is defined as the globular histone H1-domain in connection with the complete N-terminal domain and a sequence part of the C-terminal domain.

12. A therapeutic agent according to claim 8, wherein the agent is characterized by the fact that the biologically active histone H1 part is defined as the globular histone H1-domain in connection with a sequence part of the C-terminal domain as well as in connection with a sequence part of the N-terminal domain.

13. A therapeutic agent according to claim 7, wherein the agent is characterized by containing at least one human histone H1 subtype or biologically active parts thereof.

14. A therapeutic agent according to claim 13, wherein the agent is characterized by the fact that the biologically active H1.0 part is defined as the globular domain in connection with the complete N-terminal domain or sequence parts thereof, at least starting at amino acid 11, especially starting with the 16^thto the 20^thamino acid and/or in connection with the complete C-terminal domain or sequence parts thereof at least up to the 99^thto 119^thamino acid.

15. A therapeutic agent according to claim 12, wherein the agent is characterized by the fact that the biologically active H1.1 part is defined as the globular domain in connection with the complete N-terminal domain or sequence parts thereof, at least starting at amino acid 16, especially starting with the 20^thto the 26^thamino acid and/or in connection with the complete C-terminal domain or sequence parts thereof at least up to the 118^thto 138^thamino acid.

16. A therapeutic agent according to claim 13, wherein the agent is characterized by the fact that the biologically active H1.2 part is defined as the globular domain in connection with the complete N-terminal domain or sequence parts thereof, at least starting at amino acid 16, especially starting with the 20^thto the 26^thamino acid and/or in connection with the complete C-terminal domain or sequence parts thereof at least up to the 105^thto 125^thamino acid.

17. A therapeutic agent according to claim 13, wherein the agent is characterized by the fact that the biologically active H1.3 part is defined as the globular domain in connection with the complete N-terminal domain or sequence parts thereof, at least starting at amino acid 16, especially starting with the 20^thto the 26^thamino acid and/or in connection with the complete C-terminal domain or sequence parts thereof at least up to the 117^thto 137^thamino acid.

18. A therapeutic agent according to claim 13, wherein the agent is characterized by the fact that the biologically active H1.4 part is defined as the globular domain in connection with the complete N-terminal domain or sequence parts thereof, at least starting at amino acid 16, especially starting with the 20^thto the 26^thamino acid and/or in connection with the complete C-terminal domain or sequence parts thereof at least up to the 116^thto 136^thamino acid.

19. A therapeutic agent according to one or several of the previous claims, wherein the agent is characterized by the fact that it contains at least one histone and/or its biologically active sequence part, which possesses the ability to cross link the polypeptides that are aggregated in the receptor protein to larger super-structures.

20. A therapeutic agent according to one of the previous claims, wherein the agent is characterized by the fact that particular amino acids of the histones or parts thereof are substituted with amino acids of identical or similar physical properties.

21. A therapeutic agent according to claim 14, wherein the agent is characterized by the fact that the mutations K19R and K20R are introduced into the sequence of human histone H1.0. The agent can also contain further K-R mutations corresponding to the differences between histone H5 and histone H1 in birds.

22. A therapeutic agent according to one of the previous claims, wherein the agent is characterized by the fact that it resembles histone H5 (derived from chicken erythrocytes) or its biologically active part. The C-terminal tail of the latter one contains the amino acids 98-131 or 98-151.

23. A therapeutic agent according to claim 22, wherein the agent is characterized by the fact that the C-terminal sequence part is largely defined by the amino acid sequence 98 to 131 or by the amino acid sequence 98 to 151.

24. A procedure for the diagnosis of individual cancer cells having receptor proteins in their cell membrane that contain core-histones or core-like histones and/or parts thereof. The procedure utilizes proteins that can be bound to or cross-linked with the receptor protein. Such proteins are based on histones including protamines and proteins from prokaryotes and/or arche, especially histone H1, or on parts of substances that can bind to or cross-link with the receptor protein.

25. A procedure for the diagnosis of individual cancer cells having receptor proteins in their cell membrane that contain core-histones or core-like histones and/or parts thereof. The procedure utilizes antibodies that recognize the receptor proteins or structures (3-dimensional or conformational epitopes) made up of the aggregation of receptor proteins.

26. A prophylactic agent on the basis of histones or histone-like proteins (protamines, bacterial proteins from prokaryotes and/or arche), especially histone H1 or biologically active parts thereof that suppresses the generation of cancer cells expressing individual membrane-standing receptor proteins. These receptor proteins either contain different histones or histone-like polypeptides and/or parts thereof or are largely composed of them.

27. A method of using histones or covalently modified histones or functionally or structurally similar proteins or active parts thereof as marker for individual cancer cells whose individuality is determined by membrane-standing receptor proteins having the biological activities of at least one or several histones whereas the marker potential of the histones or histone-like proteins is characterized by its binding characteristics to the receptor protein.

28. The method of claim 27, wherein said marker is histone H1 or its subtypes or active parts thereof and wherein the receptor proteins are characterized by their biological activities of at least one nucleosomal core-histone or active parts thereof.

29. The method of claim 27 or 28, wherein said histones are recombinant histones or active parts thereof.

30. The method of claim 29, wherein said histones are recombinant human histones or active parts thereof.

31. A method of claim 30, wherein said histones are recombinant human histone H1 proteins or its subtypes or active parts thereof.

32. A method of claim 31, wherein said histones are recombinant histone H5 proteins or active parts thereof.

33. A method of claim 31, wherein said histones are mutated human histone H1 proteins in which all or some of the lysines have been replaced with arginine.

34. A method of one of the previous claims, wherein single amino acids of the histone H1 proteins are replaced by amino acids with similar or identical physical properties.

35. A method of one of the previous claims, wherein said markers are used for the indication of hematopoietic cancer cells.

36. A medical product for the therapy and/or prophylaxis of cancer wherein individual cancer cells are characterized by membrane-standing receptor proteins with the biological activities of one or several histones or parts thereof and wherein the therapeutic or prophylactic composition is comprised of at least a histone protein or covalently modified histones or functionally or structurally similar proteins or active parts thereof having the biological activity of binding with one or several receptor proteins in individual cancer cells and, resulting from the binding of the composition to the receptor protein, having a cytotoxic potential towards individual cancer cells.

37. A product of claim 36, wherein said composition comprises a histone H1 or a subtype of histone H1 or an active part thereof and the receptor proteins of the individual cancer cells display the biological activity of one or several core histones of the nucleosome.

38. A product of claim 36, wherein said composition comprises a recombinant human histone or a active parts thereof.

39. A product of claim 37, wherein said composition comprises a recombinant human histone H1 or its subtypes or a active parts thereof.

40. A product of claim 36, wherein said composition comprises a recombinant histone H5 or active parts thereof.

41. A product of claim 36, wherein said composition comprises a mutated human histone H1 in which all or some of the lysines have been replaced with arginine.

42. A product of claims 36 to 41, wherein single amino acids of the histone proteins are replaced by amino acids with similar or identical physical properties.

43. A product of claim 39, wherein the sequence of the recombinant human histone H1 is mutated in positions 19 and 20 by K10OR and K20R and by further mutations K→R in such a manner which corresponds to the differences between histone H1 and H5, partially or entirely.

44. A product of claims 36 to 43 for therapy and/or prophylaxis of individual cancer cells of the hematopoietic system.