EP0896584A2

EP0896584A2 - BIOLOGICALLY ACTIVE PROTEIN (COLLAGEN FRAGMENT HF-COLL-18/514cf) FOR INHIBITING THE GROWTH OF TUMOURS AND CAPILLARY PROFILERATIONS

Info

Publication number: EP0896584A2
Application number: EP97921682A
Authority: EP
Inventors: Wolf-Georg Prof.Dr.Med. Forssmann; Michael Schrader; Ludger STÄNDKER; Manfred Raida; Peter Schulz-Knappe
Original assignee: Biovision AG; FORSSMANN Wolf-Georg Prof Dr med; Haemopep Pharma GmbH
Current assignee: Digilab Biovision GmbH
Priority date: 1996-04-22
Filing date: 1997-04-22
Publication date: 1999-02-17
Also published as: JP2000511511A; WO1997040073A3; AU2766597A; WO1997040073A2; DE19615710A1

Abstract

The invention relates to a peptide from human blood, HF-COLL-18/514cf. For the purpose of diagnostic, medical and commercial use as a drug, the structure of said peptide has been identified. Isolation of said novel peptide, HF-COLL-18/514cf, proves the existence of HF-COLL-18/514cf. The molecular form of HF-COLL-18/514cf has been revealed by mass spectrometry and amino-terminal sequencing, and the fact that HF-COLL-18/514cf is a natural peptide which should be used to treat numerous diseases related to cell growth disorders, in particular of endothelial cells and vessels, and for example in cancer. HF-COLL-18/514cf can also be used commercially in a pure form or as a natural raw extract when researching new cell functions.

Description

Biologically active protein - collagen fragment HF-COLL-18 / 514cf - for inhibiting the growth of tumors and vascular growth

The present invention relates to a peptide (protein) with the ability to influence the growth of cells. The collagen fragment HF-C0LL-18 / 514cf, as well as fragments and / or derivatives derived from it, and a medicinal product containing the natural and synthetic peptides can be used for diagnostic or therapeutic purposes.

The invention relates to a method for obtaining a protein in pure or partially purified form from human body fluids, which has the ability to influence the growth of cells in a surprising manner and thus to inhibit vascular and tumor growth. A similar substance has recently been detected in the mouse (O'Reilly et al., 1997, Cell Vol.88, page 277). This substance, on the other hand, is characterized in that it can be obtained in particular from hemofiltrate or hemodialysate, which is filtered off from human blood. The substance is referred to as HF-COLL-18 / 514cf and can be used for the purpose of (1) the analysis of diseases, (2) for medical and commercial use as a drug.

The substance HF-COLL-18 / 514cf was first obtained from the hemofiltrate of kidney patients after ultrafiltration on the hemodialysis machine and was classified according to its molecular mass and 60 amino acids. the N-terminus. To produce the HF-COLL-18 / 514cf, a patented process (Forssmann, 1988; published application DE 36 33 707 AI) was refined, which was previously invented for the extraction of proteins from hemofiltrate. The fractions containing the HF-COLL-18 / 514cf can surprisingly be recognized by mass spectrometry from the molecules of a molecular weight below 20 kilodaltons obtained by this method, which are filtered off in the case of veno-venous or arterio-venous shunt connections. It was further found that, with special further processes, this substance could surprisingly be purified in such a way until a uniform protein substance was finally identified and its structure was clarified. Surprisingly, the substance is a fragment of a protein, the protein being hitherto known only at the cDNA level (Oh et al., 1994, Genomics Vol. 19, page 494). The value of this invention is characterized by the fact that this substance can be purified from the hemofiltrate previously considered worthless in order to be used as an economically usable substance.

In this way a compound was isolated, the structure of which was hitherto unknown and whose educational institution in the body is still unclear. The therapeutic and economic uses have been explored and the HF-COLL-18 / 514cf has surprisingly been recognized as an important circulating peptide of human blood.

The substance HF-COLL-18 / 514cf mentioned can be obtained by chemical synthesis and by genetic engineering production and can be used for numerous other purposes, including analysis in human blood as a pathognomonic diagnostic feature of diseases of vascular growth and growth of tumors and metastases.

The present invention thus relates to a new peptide, the HF-COLL-18 / 514cf, its production, medicaments containing it, and preparations containing it and its use therefor, and also its natural and pharmacologically contractual 1 -

are derivatives, in particular amidated, acetylated, phosphorylated and glycosylated HF-COLL-18 / 514cf derivatives and fragments of the peptide. A mean molecular weight of 18494 u daltons could be determined by mass spectrometry.

The blood peptide HF-COLL-18 / 514cf has the amino acid sequence: Val -Ala -Leu -Asn -Ser -Pro-Leu- Ser-Gly-Gly-Met-Arg-Gly-Ile -Arg -Gly- Ala-Asp-Phe -Gln-Cys-Phe-Gln-Gln-Ala-Arg-Ala -Val -Gly-Leu-Ala-Gly- Thr- Phe- Arg- Ala- Phe - Leu - Ser -Ser- Ar Q- Leu-Gin - Asr> -Leu-Tyr- Ser -Ile - Val -Arg -Arg -Ala -Asp- Arg- Ala- Ala - Val - Pro- II e- Val -Asn-Leu-Lγs -Asp- Glu-Leu-Leu- Phe-Pro-Ser-Trv-Glu-Ala -Leu-Phe-Ser-Gly-Ser-Gl--Glv- Pro-Leu -Lys-Pro-Gly-Ala-Arg-Ile-Phe-Ser-Phe-Asp- Gly-Lys-Asv-Val- Leu-Arg-His-Pro-Thr-Try-Pro-Gln-Lys-Ser-Val -TriD-His-Gly-Ser-Asp- Pro-Asn-Glv-Arg-Arg- Leu-Thr-Glu-Ser-Tyr-Cvε-Glu -Thr-Try-Arg- Thr- Glu-Ala -Pro-Ser-Ala-Thr-Gly-Gln-Ala-Ser-Ser-Leu -Leu -Gly- Gl y-Arg- Leu-Leu-Glv-Gln-Ser-Ala-Ala -Ser-Cys-His-His-Ala - Tyr-Ile-Val -Leu - Cys -Il e -Gl u -Asn - Ser - Phe -Me t- Thr -AI a- Ser

The peptide provided by the present invention, HF-COLL-18 / 514cf, is now an easily accessible drug with biological and therapeutic activity of a natural analogue of the substance found in the blood.

The present invention provides a production method for this HF-COLL-18 / 514cf and the use of the HF-COLL-18 / 514cf as a medicament for various therapeutic and diagnostic indications. For this purpose, the HF-COLL-18 / 514cf can be used as a highly pure substance or - if sufficient for the particular use - within a partially purified peptide mixture.

The peptide according to the invention, its derivatives and fragments can be produced by various methods, for example by prokaryotic or eukaryotic expression and, if appropriate, chromatographic purification. Furthermore, it can be isolated from human blood, for example by means of chromatographic methods known per se Finally, are HF-COLL-18 / 514cf or its derivatives or fragments can be prepared by conventional solid-phase and liquid-phase synthesis methods from the protected amino acids contained in the sequence given. After the protective groups have been removed, it can be purified using standard chromatography procedures.

The pharmaceutical preparation according to the invention contains HF-COLL-18 / 514cf or a physiologically tolerable salt of HF-COLL-18 / 514cf. The form and composition of the medicinal product containing the HF-COLL-18 / 514cf depends on the mode of administration. The human HF-COLL-18 / 514cf can be administered parenterally, intranasally, orally, intravenously, intramuscularly, intracutaneously, intra-thecally, locally-topically and transpulmonally. HF-COLL-18 / '514cf is preferably made up into an injection preparation, either as a solution or as a lyophilisate for dissolution immediately before use. The pharmaceutical preparation can also contain auxiliaries that are due to filling technology, contribute to the solubility, stability or sterility of the drug or increase the efficiency of absorption into the body. In particular, the use of the lyophilized form taken with mannitol in sterile ampoules for dissolution in physiological saline and / or infusion solutions for repeated single injection and / or continuous infusion in amounts of 30 micrograms to 30 milligrams of pure HF-COLL-18 / 514cf pro Therapy unit is beneficial.

The daily dose to be administered for HF-COLL-18 / 514cf depends on the indication and use of certain derivatives. At i.v./i.m. Injection is in the range from 100 to 1200 units (μg) / day, with daily subcutaneous injection preferably at 300 to 2400 units (μg) / day.

The peptide HF-COLL-18 / 514cf according to the invention is characterized in that it is also particularly suitable for long-term JPherapy for tumor diseases or other diseases which are characterized by uncontrolled vascular growth, and does not trigger an immune reaction in the case of long-term treatment. That invented The preparation according to the invention is particularly suitable for combination therapy with chemotherapy or radiation therapy or in connection with chemotherapy or radiation therapy for cancer.

The preparation according to the invention can also be used as a means of therapy and diagnosis for vascular diseases of the supporting and connective tissue, the respiratory tract, the cardiovascular system and urogenital system, the nervous system and the eye, since it can be used for the production of human-compatible antibodies are suitable for determining or influencing changes in the vascular growth in these organs.

Such antibodies can in principle be obtained by immunizing animals with the peptide according to the invention and / or its fragments or by using hybridoma technology.

The present invention also relates to a method for treating patients who need HF-COLL-18 / 514cf or its derivatives or fragments by administering therapeutic amounts of HF-COLL-18 / 514cf. Patients who suffer from excessive production of HF-COLL-18 / 514cf or its derivatives or fragments require the administration of therapeutic amounts of an antagonist / inhibitor of HF-COLL-18 / 514cf.

The medicament according to the invention is suitable for the treatment of diseases of the human organism, in particular in connection with vascular growths, cancer diseases, diseases involving the cardiovascular and nervous systems, diseases involving the intugement and the sensory organs, in particular the eye.

According to the invention, the use of the peptide or its derivatives, the fragments or the antibody according to the invention for the manufacture of a medicament for the treatment of disorders in inflammatory processes, disturbed inflammatory reactions, proliferation and maturation disorders of the hematopoietic system, system diseases in the case of overproduction or lack of HF COLL-18 / 514cf, in particular in the case of antibodies formed, for example, by use against this or for the use of HF-COLL-18 / 514cf for substitution therapy, chronic diseases, partly associated with the diseases mentioned, by being in a suitable form for the treatment for the electrolyte - Effect is used in tumor and vascular diseases.

The medicament according to the invention is suitable for the treatment of acute diseases of the type mentioned above, in that it is used in a corresponding form for the treatment in the intensive care of these diseases.

A further use of the peptide according to the invention, its fragments or the antibody according to the invention is used to diagnose diseases by producing specific antibodies against synthetic parts or the entire peptide or its derivatives and its fragments and e.g. the blood concentration of the HF-COLL-18 / 514cf is measured by immunoassays.

A diagnostic agent containing the peptide according to the invention, its fragments or antibodies according to the invention for test systems for checking tissue, plasma, urine and cerebrospinal fluid levels of this substance are thus also the subject of the invention. The diagnostic agent according to the invention is particularly suitable as a marker for certain cancers and for functional disorders of blood vessels, bone marrow, lymph organs, the gastrointestinal tract, the immune system and inflammatory and neoplastic processes.

The invention will be closer he explained ^¬ by the following examples. Example 1: Isolation and characterization of circulating HF-COLL-18 / 514cf from human hemofiltrate

Hemofiltrate, which is obtained in large quantities in the treatment of kidney insufficient patients and all plasma components up to a molecular size of about 20, was used as the starting material. Contains 000 daltons.

I. Obtaining the raw peptide material

The hemofiltrate was obtained by means of a Sartorius hemofiltration system using cellulose triacetate filters with an exclusion size of 20,000 daltons (type SM 40042, Sartoriuε, Göttingen, FRG). The filtrate came from kidney insufficiency patients who were in a stable metabolic state due to long-term hemofiltration and was protected against proteolytic degradation by acidification and cooling to 4 ° C immediately after it was obtained. In four extractions with a cation exchange column (TSK SP 650 (M), Merck, Darmstadt, DE), 2860 l of hemofiltrate were processed. 93% of the pooled extracts were successively eluted on the column material mentioned above using different buffers with different pH values. The crude fractions were then freeze-dried.

II. Preparative RP chromatography

500 mg of 2200 mg of the last crude fraction were roughly separated for hydrophobicity by preparative RP chromatography. Fractions were collected from a PrepPak cartridge with the dimensions 47 x 300 mm from Waters. Fraction 31 was used for further purification.

Device: BioCad HPLC (Perseptive Biosystems, Freibύrg, DE)

Column: Waters PrepPak Cartridge 47 x 300 mm

Material: Vydac, 300 Ä, 15 - 20 μm

Eluent A: water with 10MM HC1 Eluent B: methanol with 10 mM HC1 gradient: 0 - 50% eluent B 28.57 min 50 - 95% eluent B 61.43 min 95% eluent B 5.71 min flow rate: 35 ml / min fractions: 50 ml or 1 , 43 min detection: 230 nm and 280 nm

III. First analytical RP-HPLC

UV absorption during analytical RP chromatography of fraction 31, which was obtained from the separation in Figure 1. A further separation was achieved in a gradient on a Vydac column (10 x 250 mm steel jacket, material: RP C18, 300 Å, 5 μm). The eluents were water with 0.1 vol% trifluoroacetic acid and acetonitrile with 0.1 vol% trifluoroacetic acid.

Device: Kontron HPLC system Column: Vydac, steel jacket 10 x 250 mm Material Vydac RP-C18, 300 Ä, 5 μm Eluent A water with 0.1 vol% trifluoroacetic acid Eluent B acetonitrile with 0.1 vol% trifluoroacetic acid gradient 0 - 60 % Eluent B 50 min

60 - 80% eluent B 5 min

80 - 0% eluent B 5 min

Flow rate: 2 ml / min fractions 2 ml or 1 min detection: 230 nm

IV. Discovery of the molecular mass of the HF-COLL-18 / 514cf using MALDI-TOF mass spectrometry

Using a MALDI mass spectrometer RBT II (Vestec / Per-Septive, Houston, Texas, USA), mass spectra were obtained from the native, purified HF-COLL-18 / 514cf from the preparation in step III with alpha-cyano-4-hydroxycinnamic acid as a matrix measured. In the Fractions 45 and 46 showed peaks of the single, double and triple protonated molecule, with a molecular mass of about 18500 u. Various minor components were also visible.

V. Second analytical RP-HPLC

In a final analytical RP chromatography of pooled fractions 45 and 46, which were obtained from the separation in step III, 25 highly purified HF-COLL-18 / 514cf could be isolated in the fraction.

Device: Kontron HPLC (Kontron, Munich, DE) Column: YMC, steel jacket 4.6 x 250 mm Material YMC RP-C18, 300 Ä, 5 μm eluent A water with 0.1 vol% trifluoroacetic acid eluent B 80% acetonitrile, 20 % Water

(V: V) with 0.1 vol% trifluoroacetic acid

Gradient 0 - 30% eluent B 5 min

30 - 80% eluent B 150 min

80 - 100% eluent B 5 min

100% eluent B 5 min

Flow rate: 0.6 ml / min Fractions: manually collected Detection: 230 nm and 280 nm

VI. Purity determination using capillary zone electrophoresis

5 ul of fraction 25 were used directly for measurement in capillary zone electrophoresis. The electropherogram shows only one peak and no further peaks of secondary components. This result shows that a high-purity HF-COLL-18 / 514cf was present in the final cleaning stage.

Device: P / ACE System 2000, Beckman Instruments GmbH,

Munich, DE Capillary: Uncoated Fuεed Silica, 500 mm x 75 μm ID Buffer: 100 mM sodium phosphate pH 2.5

0.02% hydroxypropylmethyl cellulose

Temperature: 25 ° C

Injection: 20 see corresponds to 120 nl

Running time: 25 minutes

Current: 80 μA, constant

Detection: 200 nm

VII. Determination of the molecular mass of HF-COLL-18 / 514cf

Using MALDI-TOF mass spectrometry on a Vestec BT II of the native, purified HF-COLL-18 / 514cf from fraction 25 in step V, spectra could be obtained on two matrices (alpha-cyano-4-hydroxy-cinnamic acid and 2,5-dihydroxybenzoic acid) be preserved. Eε show peaks of the single, double and triple protonated molecule. The molecular mass is determined to be 18507 u ± 20 u. Minor components are not visible.

For a more precise determination of the molecular mass of the native, purified HF-COLL-18 / 514cf, a mass spectrum was additionally measured from fraction 25 of step V using an electrospray mass spectrometer (Sciex API III, Perkin-Elmer, Langen, DE). Peaks of the eight to eleven times protonated molecule are shown. The average molecular mass of HF-COLL-18 / 514cf is determined here at 18494 u + 3 u, the theoretical value is 18496 u (see VIII).

VIII. Determination of the amino terminal amino acid sequence

The first 60 amino acids were determined by means of automated Edman sequencing with an ABI 494 gas-phase amino acid sequencer (Applied Biosystems, Perkin-Elmer, Weiterstadt, DE). No amino acid was detected at the 21st position (Xxx), as is customary for cysteine.

Val -Ala -Leu-Asn-Ser-Pro-Leu -Ser-Gl y-Gly-Met -Arg-Gl y- Ile-Arg-Gl y- Ala -Asp-Phe-Gln-Xxx-Phe-Gln-Gln -Ala -Arg-Ala -Val -Gl v-Leu -Al a -Gl y- Thr- Phe-Arg-Ala-Phe-Leu-Ser-Ser- Arg- Leu-Gin- Asu-Leu -Tyr -Ser- Ile - Val -Arg-Arg-Ala -Asp-Arg-Ala -Ala -Val - Pro-Il e-Val It was thus established that it is a fragment of collagen alpha 1 (XVIII), whereby this protein is only known at the cDNA level (Oh et al., 1994, Genomics Vol. 19, page 494). The fragment begins at position 514 of the protein precursor and the molecular mass shows that it ends at the penultimate position of the precursor with the amino acid serine, that is to say C-terminally shortened by a lysine.

Example 2: Investigation of the biological effectiveness of HF-COLL-18 / 514cf

Using the method shown in Example 1, a larger amount of material of more than 0.1 mg HF-COLL-18 / 514cf was isolated from human hemofiltrate. The high-purity HF-COLL-18 / 514cf was used to determine the biological function in endothelial cell proliferation assays. For this assay, bovine capillary endothelial cells from the adrenal cortex of freshly slaughtered calves were cultured as described in the literature (Folkman et al., 1979, Proc. Natl. Acad. Sci. Vol. 76, page 5217).

The proliferation assay was carried out as described in the literature (O'Reilly et al., 1997, Cell Vol.88, page 277). For this purpose, the bovine capillary endothelial cells were washed with PBS (phosphate buffer with sodium chloride, pH 7.4) and suspended in a 0.05% trypsin solution. A cell suspension with 25000 cells per ml in DMEM medium, containing 10% FCS (fetal calf serum) and 1% GPS (glutamine-penicillin-streptomycin) was placed in gelatin-coated 24-hole plates (0.5 ml per Hole) at 37 ° C and 10% CO ₂ .

After 24 h the medium was replaced by 0.5 ml DMEM medium containing 5% FCS and 1% GPS as well as different concentrations (from 0 to 1000 ng / ml final concentration) of the isolated, high-purity HF-COLL-18 / 514cf , replaced. After another 30 Minutes of incubation were added to the batches bFGF (basic fibroblast growth factor) in a final concentration of 1 ng / ml. After 72 h the number of cells in the batches was determined by crystal violet staining of the cells and measurement of the absorption at 600 nm. The HF-COLL-18 / 514cf added to the bovine capillary endothelial cells inhibited the proliferation of these cells stimulated with bFGF in a concentration-dependent manner. Half-maximal inhibition of proliferation in this assay was achieved at a concentration of 200 ng / ml HF-COLL-18 / 514cf.

In order to investigate the specificity of the spectrum of activity of HF-COLL-18 / 514cf and thus other possible biological functions, proliferation assays were carried out with non-endothelial cells. In tests with fibroblast cell lines, namely NIH 3T3 cells and LMTK cells, HF-COLL-18 / 514cf showed no significant activity and therefore no antiproliferative activity.

SEQUENCE LOG

(1. GENERAL INFORMATION:

(i) APPLICANT:

(A) NAME: Wolf-Georg Forssmann

(B) STREET: Feodor-Lynen-Str. 31

(C) LOCATION: Hanover

(E) COUNTRY: Germany

(F) POSTAL NUMBER: 30625

(ii) NAME OF THE INVENTION: Biologically active protein collagen fragment HF-COLL-18 / 514cf - for inhibiting the growth of tumors and vascular growth liii) NUMBER OF SEQUENCES: 1

(iv) COMPUTER READABLE VERSION:

(A) DISK: Floppy disk

(B) COMPUTER: IBM PC compatible

(C) OPERATING SYSTEM: PC-DOS / MS-DOS

(D) SOFTWARE: Patentin Release # 1.0, Version # 1.30 (EPA)

(2) INFORMATION ON SEQ ID NO: 1:

(i) SEQUENCE LABEL:

(A) LENGTH: 170 amino acids

(B) TYPE: amino acid

(C) STRAND FORM: Single strand

(D) TOPOLOGY: not known

(ii) MOLECULE TYPE: Peptide (Üi) HYPOTHETICAL: NO

(xi) SEQUENCE DESCRIPTION: SEQ ID NO: 1:

Val Ala Leu Asn Ser Pro Leu Ser Gly Gly Met Arg Gly Ile Arg Gly 1 5 10 15

Ala Asp Phe Gin Cys Phe Gin Gin Ala Arg Ala Val Gly Leu Ala Gly 20 25 30

Thr Phe Arg Ala Phe Leu Ser Ser Arg Leu Gin Asp Leu Tyr Ser Ile 35 40 45

Val Arg Arg Ala Asp Arg Ala Ala Val Pro Ile Val Asn Leu Lys Asp 50 55 60

Glu Leu Leu Phe Pro Ser Trp Glu Ala Leu Phe Ser Gly Ser Glu Gly 65 70 75 80

Pro Leu Lys Pro Gly Ala Arg Ile Phe Ser Phe Asp Gly Lys Asp Val 85 90 95

Leu Arg His Pro Thr Trp Pro Gin Lys Ser Val Trp His Gly Ser Asp 100 105 110 Pro Asn Gly Arg Arg Leu Thr Glu Ser Tyr Cys Glu Thr Trp Arg Thr 115 120 125

Glu Ala Pro Ser Ala Thr Gly Gin Ala Ser Ser Leu Leu Gly Gly Arg 130 135 140

Leu Leu Gly Gin Ser Ala Ala Ser Cys His His Ala Tyr Ile Val Leu 145 150 155 160

Cys Ile Glu Asn Ser Phe Met Thr Ala Ser

165 170

Claims

Expectations

Peptide with the amino acid sequence

Val -Ala - Leu- Asn- Ser -Pro-Leu- Ser-Glγ-Gly-Met- Arg-Gly-Ile- Arg-

Gly-Ala -Asv-Phe-Gln -Cvs-Phe-Gln-Gln-Ala-Arg-Ala-Val -Gly-Leu-

Ala-Gly-Thr-Phe-Arg-Ala-Phe-Leu-Ser-Ser-Arg-Leu-Gln-Asp-Leu-

Tγr-Ser-Ile-Val -Arg-Arg-Ala-Asp-Arg-Ala-Ala -Val -Pro-Ile-Val -

Aεn-Leu-Lγs-Asp-Glu-Leu-Leu-Phe-Pro-Ser-Trp-Glu-Ala-Leu-Phe-

Ser-Gl y-Ser-Glu-Glγ-Pro-Leu-Lvs-Pro-Glv-Ala-Arg-Ile-Phe-Ser-

Phe-Asp-Gly-Lγs-Asp-Val -Leu-Arg-His-Pro-Thr-TrO-Pro-Gln-Lvs-

Ser-Val - Trv-His-Gly-Ser-Asp-Pro-Asn-Gly-Arg-Arg-Leu-Thr-Glu-

Ser-Tyr-Cyε-Glu-Thr-Trp-Arg-Thr-Glu-Ala-Pro-Ser-Ala-Thr-Gly-

Gln-Ala-Ser-Ser-Leu-Leu-Gly-Glγ-Arg-Leu-Leu-Gly-Gln-Ser-Ala-

Ala-Ser-Cys-His-His-Ala-Tyr-Ile-Val -Leu-Cys-Ile-Glu-Asn-Ser-

Phe -Met - Thr -Ala -Ser (HF-COLL-18 / 514cf) as well as its natural and pharmacologically compatible

Derivatives, especially amidated, acetylated, phosphorylated and glycosylated derivatives.

Fragments of the peptide according to claim 1 which are pharmacologically active.

A process for the preparation of the peptide according to claim 1 and / or its fragments according to claim 2, characterized gekenn¬ characterized in that it is produced via a prokaryotic or a eukaryotic expression.

A method for producing the peptide according to claim 1 and / or its fragments according to claim 2, characterized in that it is isolated from human blood using a chromatographic method. 5. A process for the preparation of the peptide or its derivatives according to claim 2 and its fragments according to claim 2, characterized in that the peptide or its derivatives or fragments by conventional methods of solid phase and liquid phase synthesis from the protected amino acids , which are contained in the output sequence, are produced, deblocked and cleaned using known chromatographic methods.

6. Medicaments containing the peptide according to claim 1 or its fragments according to claim 2 as an active ingredient in addition to usual auxiliaries and additives.

7. Medicament according to claim 6 for oral, parenteral, intravenous, intramuscular, intracutaneous, intrathecal, intranasal and topical local application and as an aerosol for transpulmonary application.

8. Antibodies obtainable by immunizing animals with a peptide according to claim 1 and / or fragments according to claim 2 and / or by using the hybridoma technology.

9. A method for the treatment of patients who need HF-COLL-18 / 514cf or its derivatives or fragments according to claim 1 by administration of therapeutic amounts of HF-COLL-18 / 514cf.

10. A method of treating patients who require inhibition of HF-COLL-18 / 514cf or its derivatives or fragments according to claim 1 or 2 by administering therapeutic amounts of an antagonist / inhibitor of HF-COLL-18 / 514cf.

11. Use of the medicament according to claims 6 or 7 for the treatment of diseases of the human organism, in particular in connection with vascular growths. 12. Use of the medicament according to claims 6 or 7 for the treatment of diseases of the human organism, in particular cancer.

13. Use of the medicament according to claims 6 or 7 for the treatment of diseases of the human organism, in particular with participation of the cardiovascular and nervous system.

14. Use of the medicament according to claims 6 or 7 for the treatment of diseases of the human organism, in particular with the participation of the intugement and the sensory organs, in particular the eye.

15. Use of the peptide or its derivatives according to claim 1, the fragments according to claim 2 or the antibody according to claim 8 for the manufacture of a medicament for the treatment of disorders in inflammatory processes, disturbed inflammatory reactions, proliferation and maturation disorders of the hematopoietic system.

16. Use of the medicament according to claims 6 or 7 or the antibody according to claim 8 for the treatment of systemic diseases in the case of overproduction or deficiency of HF-COLL-18 / 514cf, in particular in e.g. by application against this antibody formed or by using HF-COLL-18 / 514cf for substitution therapy.

17. Use of the medicament according to claims 6 or 7 for the treatment of chronic diseases, partly socialized with the diseases mentioned in claims 11 to 16, by using it in a suitable form for the treatment for the electrolyte effect in tumor and vascular diseases.

18. Use of the medicament according to claims 6 or 7 or of the antibody according to claim 8 for the treatment of acute Diseases, according to claims 11 to 16, in that it is used in a suitable form for the treatment in the intensive care of these diseases.

19. Use of the medicament according to claims 6 or 7 or the antibody according to claim 8 for the diagnosis of diseases, in particular according to any one of claims 11 to 16, by producing specific antibodies against synthetic parts or the entire peptide or its derivatives and its fragments and the blood concentration HF-COLL-18 / 514cf is measured by immunoassays.

20. Diagnostic agent containing the peptide according to claim 1, fragments according to claim 2 or antibodies according to claim 8 for test systems for the control of tissue, plasma, urine and cerebrospinal fluid levels of this substance.

21. Diagnostic agent according to claim 20 as a marker for certain cancers and for functional disorders of blood vessels, bone marrow, lymph organs, the gastrointestinal tract, the immune system and inflammatory and neoplastic processes.