Title of Invention

FACTOR VII OR VIIa-LIKE MOLECULES

Abstract

ABSTRACT IN/PCT/2002/01373/CHE "FACTOR VII OR VIIa-LIKE MOLECULES" The present invention relates to novel factor VII (FVII) or Factor Vila (FVIIa) polypeptide conjugates, to their preparation and use in therapy, in particular for the treatment of variety of coagulation-related disorders. These novel polypeptide conjugates comprise at least one non-polypeptide moiety covalently attached to a polypeptide, wherein the amino acid sequence of the polypeptide differs from that of wild-type FVII or FVIIa in that at least one amino acid residue comprising an attachment group for said non polypeptide moiety has been introduced or removed. The conjugates of the present invention have one or more improved properties as compared to commercially available rFVIIa, including increased functional in vivo half-life and/or increased plasma half-life, and/or increased bioavailability and/or reduced sensitivity to proteolysis degradation.

Full Text

FIELD OF 1 HE IN VEN1 lUN The present invention relates to novel Factor VH (FVII) or Factor VHa (FVILa) polypeptide conjugates, to their preparation and use in therapy, in particular for the treatment of a variety of coagulation-related disorders. BACKGROUND OF THE ENTION Blood coagulation is a process consisting of a complex interaction of various blood components (or factors) that eventually gives rise to a fibrin clot. Generally, the blood components participating in what has been referred to as the coagulation "cascade" are proenzymes or zymogens, i.e. enzymatically inactive proteins that are converted into an active form by the action of an activator. One of tiles coagulation factors is FVII. Fan is a vitamin K-dependent plasma protein synthesized in the liver and secreted into the blood as a single-chain glycoprotein with a molecular weight of 53 kDa (Broze & Majerus, J. Biol. Chem 1980; 255:1242-1247). The FVII zymogene is converted into an activated form (FVIIa) by proteolytic cleavage at a single site, R152-I153, resulting in two chains linked by a single disulfide bridge. FVIIa in complex with tissue factor (FVHa complex) is able to convert both factor DC and factor X into their activated forms, followed by reactions leading to rapid thrombin production and fibrin formation (0sterud & Rapaport, Proc Natl Acadia Sci USA 1977; 74:5260-5264). Fun undergoes post-translational modifications, including vitamin K-dependent carbox3dation resulting in ten Y-carboxyglutamic acid residues in the N-terminal region of the molecule. Thus, residue number 6, 7,14,16, 19, 20, 25,26, 29 and 35 shown in SEQ ID N0:1 are Y-carboxyglutamic acids residues in the Gla domain important for FVH activity. Other post-tiranslational modifications include sugar moiety attachment at two naturally occurring N-glycosylation sites at position 145 and 322, respectively, and at two naturally occurring O-glycosylation sites at position 52 and 60, respectively. The gene coding for human FVH (hFVII) has been mapped to cliromosome 13 at q34-gter 9 (de Grouchy et al.. Hum Genet 19S4; 66:230-233). It contains nine exons and spans 12.8 Kb (O'Hara et al., Proc Nad Acad Sci USA 1987; 84:5158-5162). The gene organisation and protein structure of FVH are similar to those of other vitamin K-dependent procoagulant proteins, with exons la and lb encoding for signal sequence; exon 2 the propeptide and Gla - domain; exon 3 a short hydrophobic region; exons 4 and 5 the epidermal growth factor-like domains; and exon 6 througl^ 8 the serine protease catalytic domain (Yoshitake et al., Biochemistry 1985; 24: 3736-3750). Reports exist on experimental three-dimensional structures of hFVIIa (Hke et al., PNAS. U.S.A., 1999; 96:8925-30 and KembaU-Cook et al., J.StractBiol, 1999; 127:213-223), of hFVIIa in complex with soluble tissue factor using X-ray crystallographic methods (Banner et al., Nature, 1996; 380:41 and Zhang et al., J.Mol.Biol, 1999; 285: 2089), and of smaller fragments of hFVH (Muranyi et al., Biochemistry, 1998; 37:10605 and Kao et al., Biochemistry, 1999; 38:7097). Relatively few protein-engineered variants of FVH have been reported (Dickinson & Ruf, J Bio Chem, 1997;272:19875-19879, Kemball-Cook et al., J Biol Chem, 1998; 273:8516-8521, Bharadwaj et al., J Biol Chem, 1996; 271:30685-30691, Ruf et al., Biochemistry, 1999; 38:1957-1966). Reports exist on expression of FVII in BHK or other mammalian cells (W092/15686, W091/11514 and WO88/10295) and co-expression of FVn and kex2 endoprotease in eulcaryotic cells (WO 00/28065). Commercial preparations of human recombinant FVIIa are sold as NovoSeven®. NovoSeven® is indicated for the treatment of bleeding episodes in hemophilia A or B patients. NovoSeven® is the only rPVUa for effective and reliable treatment of bleeding episodes available on the market. An inactive form of FVII in which argjnine 152 and/or isoleucine 153 is/are modified has been reported in W091/1154. These amino acids are located at the activation site. WO 96/12800 describes inactivation of FVIIa by a serine proteinase inhibitor; inactivation by carbamylation of FVIIa at the a-amino acid group 1153 has been described by Petersen et al., Eur J Biochem, 1999;261:124-129. The inactivated form is capable of competing with wild-type FVn or FVUa for binding to tissue factor and inhibiting clotting activity. The inactivated form of FVUa is suggested to be used for treatment of patients being in hypercoagulable states, such as patients with sepsis, in risk of myocardial infarction or of thrombotic stroke. A circulating rFVUa half-life of 2.3 hours was reported in "Summary Basis for Approval for NovoSeven®", FDA reference number 96-0597. Relatively high doses and frequent administration are necessary to reach and sustain the desired therapeutic or prophylactic effect. As a consequence adequate dose regulation is difficult to obtain and the need of frequent intravenous administrations imposes restrictions on the patient's way of living. Another problem in current rFVIIa ti-eatment is the relative instability of the molecule with respect to proteolytic degradation. Proteolytic degradation is a major obstacle for obtaining a preparation in solution as opposed to a lyophilised product. The advantage of obtaining a stable soluble preparation lies in easier handling for the patient, and, in the case of emergencies, quicker action, which potentially can become life saving. Attempts to prevent proteolytic degradation by site directed mutagenesis at major proteolytic sites have been disclosed in WO88/10295. A molecule with a longer circulation half-life would decrease the number of necessary administrations. Given the association of current FVIIa product with frequent injections, and the potential for obtaining more optimal therapeutic FVIIa levels with concomitant enhanced therapeutic effect, there is a clear need for improved FVn or FVIIa-like molecules. One way to increase the circulation half-life of a protein is to ensure that renal cleai-ance of the protein is reduced. This may be achieved by conjugating the protein to a chemical moiety, which is capable of conferring reduced renal clearance to the protein. Furthermore, attachment of a chemical moiety to the protein or substitution of amino acids exposed to proteolysis may effectively block a proteolytic enzyme from contact leading to proteolytic degradation of the protein. Polyethylene glycol (PEG) is one such chenaical moiety that has been used in the preparation of therapeutic protein products. W098/32466 suggests that FVn, among many other proteins, may be PEGylated but does not contain any further information in this respect. BRIEF DISCLOSURE OF THE INVENTION This application discloses impro^'ed FVn and FVUa molecules, in particular recombinant hFVH and hFVILa molecules, providing one or more of the aforementioned desired benefits. Thus, the conjugate of the present invention has one or more improved properties as compared to commercially available rFVIIa, including increased functional in vivo half-life and/or increased plasma half-life, and/or increased bioavailability and/or reduced sensitivity to proteolytic degradation. Consequently, medical treatment with a conjugate of the invention offers a number of advantages over the currently available rFVIIa compound, such as longer duration between injections. Accordingly, in a first aspect the invention relates to a conjugate comprising at least one non-polypeptide moiety covalentiy attached to a polypeptide, wherein the amino acid sequence of the pol}'peptide differs from that of wild-type FVH or FVUa shov^'n in SEQ ID ' N0:1 in that at least one amino acid residue comprising an attachment group for said non-polypeptide moiety has been introduced or removed. In another aspect the invention relates to a polypeptide, wherein the amino acid sequence of the polypeptide differs from that of wild-type FVH or hFVIIa shown in SEQ ID N0:1 in that at least one amino acid residue comprising an attachment group for a non-polypeptide moiety has been introduced or removed. Such novel FVH polypeptides are contemplated to be useful as such for therapeutic, diagnostic or other purposes, but find particular interest as intermediate products for the preparation of a conjugate of the invention. In further aspects the invention relates to: a nucleotide sequence encoding the polypeptide of the invention or the polypeptide part of the conjugate of the invention; an expression vector harbouring the nucleotide sequence of the invention; a host cell comprising the nucleotide sequence of the invention or the expression vector of the invention. In still further aspects the invention relates to pharmaceutical compositions comprising the conjugate of the invention as well as to methods for preparing and using such conjugates. DETAILED DISCLOSURE OF THE INVENTION Definitions The term "conjugate" (or interchangeably "conjugated polypeptide") is intended to indicate a heterogeneous (in the sense of composite or chimeric) molecule formed by the covalent attachment of one or more polypeptide(s) to one or more non-polypeptide moieties such as polymer molecules, lipophilic compounds, sugar moieties or organic derivatizing agents. Preferably, the conjugate is soluble at relevant concentrations and conditions, i.e. soluble in physiological fluids such as blood. Examples of conjugated polypeptides of the invention include glycosylated and/or PEGylated polypeptides. The term "covalent attachment" means that the polypeptide and the non-polypeptide moiety are either directly covalently joined to one another, or else are indirectly covalently joined to one another through an intervening moiety or moieties, such as a bridge, spacer, or linkage moiety or moieties. The term "non-conjugated polypeptide" may be used about the polypeptide part of the conjugate. When used herein, the term "non-polypeptide moiety" means a molecule that is capable of conjugating to an attachment group of the polypeptide of the invention. Preferred examples of such molecules include poljoner molecules, sugar moieties, lipophilic compounds, or organic derivatizing agents. When used in the context of a conjugate of the invention it will be understood that the non-polypeptide moiety is linlced to the polypeptide part of the conjugate through an attachment group of the pol5'peptide. As explained above, the non-polypeptide moiety may be directly covalently joined to the attachment group or it may be indirectly covalently joined to the attachment group through an intervening moiety or moieties, such as a bridge, spacer, or linkage moiety or moieties. The "polj^er molecule" is a molecule formed by covalent linkage of t\\'o or more monomers, wherein none of the monomers is an amino acid residue, except where the polymer is human albumin or another abundant plasma protein. The term "polymer" may be used interchangeably with the term "polymer molecule". The term is intended to cover carbohydrate molecules attached by in vitro glycosylation, i.e. a synthetic glycosylation performed in vitro normally involving covalently hnlcing a carbohydrate molecule to an attachment group of the polypeptide, optionally using a cross-linlcing agent. Carbohydrate molecules attached by in vivo glycosylation, such as N- or O-glycosylation (as further described below) are referred to herein as a "sugar moiety". Except where the number of non-polypeptide moieties, such as polymer molecule(s) or sugar moieties in the conjugate is expressly indicated every reference to "a non-polypeptide moiety" contained in a conjugate or otherwise used in the present invention shall be a reference to one or more non-polypeptide moieties, such as polymer molecule(s) or sugar moieties. The term "attachment group" is intended to indicate a functional group of the polypeptide, in particulai' of an amino acid residue thereof or a carbohydrate moiety, capable of attaching a non-polypeptide moiety such as a polj'mer molecule, a lipophilic molecule, a sugar moiety or an organic derivatizing agent. Useful attachment gi'oups and their matching non-polypeptide moieties are apparent from the table below. Attaclmient Amino acid Examples of non- Conjugation Reference group polypeptide moiety method/-Activated PEG -NHz N-terminal, Polymer, e.g. PEG, mPEG-SPA Shearwater Inc. Lys with amide or imine group Tresylated mPEG Delgado et al, critical reviews in Therapeutic Drug Carrier Systems 9(3,4):249-304 (1992) -COOH C-tenninal, Asp, Glu Polymer, e.g. PEG, with ester or amide group Carbohydrate moiety mPEG-Hz In vitro coupling Shearwater Inc. -SH Cys Polymer, e.g. PEG, PEG-vinylsul- Shearwater Inc; with disulfide, phone Delgado et al, critical maleimide or vinyl sulfone group PEG-maleimide reviews in Therapeutic Drug Carrier Systems t moiety (1992) -OH Ser, Tlir, Lys, OH- Sugar moiety PEG with ester, ether, carbamate, carbonate In vivo 0-linked glycosylation -CONH2 Asn as part of an N-glycosyla-tion site Sugar moiety Polymer, e.g. PEG In vivo N-glycosylation Aromatic Phe, Tyr, Carbohydrate In vitro coupling residue Trp moiety -1 -C0NH2 Gk Carbohydrate moiety In vitro coupling Yan and Wold, Biochemistry, 19S4, Jul 31; 23(16): 3759-65 Aldehyde Oxidized Polymer, e.g. PEG, PEGylation Andresz et al., 1978, Ketone oligosaccharide PEG-hydrazide Malaomol. Chem. 179:301, WO 92/16555, WO 00/23114 Guanidino Arg Carbohydrate moiety In vitro coupling Lundblad and Noyes, Chimical Reagents for Protein Modification, CRC Press Inc., Florida, USA Imidazole His Carbohydrate In vitro coupling As for guanidine ring moiety For in vivo N-glycosylation, the term "attachment gi"oup" is used in an unconventional way to indicate the amino acid residues constituting a N-glycosylation site (with the sequence N-X-S/T/C, wherein X is any amino acid residue except proline, N is asparagine and S/T/C is either serine, threonine or cysteine, preferably serine or threonine, and most preferably threonine). Although the asparagine residue of the N-glycosylation site is the one to which the sugar moiety is attached during glycosylation, such attachment cannot be achieved unless the other amino acid residues of the N-glycosylation site is present. Accordingly, when the non-pol}'peptide moiety is a sugar moiety and the conjugation is to be achieved by N-glycosylation, the term "amino acid residue comprising an attachment group for the non-polypeptide moiety" as used in connection with alterations of the amino acid sequence of the polypeptide of interest is to be understood as meaning that one or more amino acid residues constituting an N-glycosylation site are to be altered in such a manner that either a functional N-glycosylation site is introduced into the amino acid sequence or removed from said sequence. In the present application, amino acid names and atom names (e.g. CA, CB, CD, CG, SG, NZ, N, O, C, etc) are used as denned by the Protein DataBank (PDB) (www.pdb.org) based on the lUPAC nomenclature (TUPAC Nomenclature and Symbolism for Amino Acids and Peptides (residue names, atom names, etc.), Eur. J. Biochem., 138, 9-37 (1984) together with their corrections in Eur. J. Biochem.., 152,1 (1985)). The tenn "amino acid residue" is intended to indicate an amino acid residue contained in the group consisting of alanine (Ala or A), cysteine (Cys or C), aspartic acid (Asp or D), glutamic acid (Glu or E), phenylalanine (Phe or F), glycine (Gly or G), histidine (His or H), isoleucine (Be or I), lysine (Lys or K), leucine (Leu or L), methionine (Met or M), asparagine (Asn or N), proline (Pro or P), glutamine (Gin or Q), arginine (Arg or R), serine (Ser or S), threonine (Thr or T), valine (Val or V), tryptophan (Trp or W), and tyrosine (Tyr or Y) residues. The tenninology used for identifying amino acid positions is illustrated as follows: G124 indicates that position 124 is occupied by a glycine residue in the amino acid sequence shown in SEQ ID NO: 1. G124R indicates that the glycine residue of position 124 has been substituted with an arginine residue. Alternative substitutions are indicated with a "/", e.g. EC32D/E means an amino acid sequence in which lysine in position 32 is substituted with either aspartic acid or glutamic acid. Multiple substitutions are indicated with a "+", e.g. K.143N+N145S/T means an amino acid sequence which comprises a substitution of the lysine residue in position 143 with an asparagine residue and a substitution of the asparagine residue I.XA fci/V/UAUW^l.* a."T*^ ry A.*rt.A 1.C U^>rxJLJ. residue, such as insertion of an alanine residue after G124 is indicated by G124GA. A deletion of an amino acid residue is indicated by an asterix. For example, deletion of a glycine in position 124 is indicated by G124*. Unless otherwise indicated, the numbering of amino acid residues made herein is made relative to the amino acid sequence of wild-type FVn/FVUa shown in SEQ ID NO: 1. The term "differs from" as used in coimection with specific mutations is intended to allow for additional differences being present apart from the specified amino acid difference. For instance, in addition to the removal and/or introduction of amino acid residues comprising an attachment group for the non-polypeptide moiety the FVn or FVHa polypeptide may comprise other substitutions that are not related to introduction and/or removal of such amino acid residues. Thus, in addition to the amino acid alterations disclosed herein aimed at removing and/or introducing attachment sites for the non-polypeptide moiety, it will be understood that the amino acid sequence of the polypeptide of the invention may, if desired, contain other alterations that need not be related to introduction or removal of attachment sites, i.e. other substitutions, insertions or deletions. These may, for example, include truncation of the N- and/or C-terminus by one or more amino acid residues, or addition of one or more extra residues at the N- and/or C-terminus, e.g. addition of a methionine residue at the N-terminus as well as "conservative amino acid substitutions", i.e. substitutions performed within groups of amino acids with similar characteristics, e.g. small amino acids, acidic amino acids, polar amino acids, basic amino acids, hydrophobic amino acids and aromatic amino acids. Preferred substitutions in the present invention may in particular be selected from the conservative substitution groups listed in the table below. 1 Alanine (A) Glycine (G) Serine (S) Threonine (T) 2 Aspartic acid (D) Glutamic acid (E) 3 Asparagine (N) Glutamine (Q) 4 Arginine (R) Histidine (H) Lysine (K) 5 Isoleucine (I) Leucine (L) Methionine (M) Valine (V) 6 Phenylalanine (F) Tyrosine (Y) Tryptophan (W) .,vv- The terms "mutation" and "substitution" are used interchangeably herein. The term "nucleotide sequence" is intended to indicate a consecutive stretch of two or more nucleotide molecules. The nucleotide sequence may be of genomic, cDNA, RNA, semisj'nthetic, synthetic origin, or any combinations thereof. The term "polymerase chain reaction" or "PCR" generally refers to a metliod for amplification of a desired nucleotide sequence in vitro, as described, for example, in US 4,683,195. In general, the PCR method involves repeated cycles of primer extension sjoithesis, using oligonucleotide primers capable of hybridising preferentially to a template nucleic acid. "Cell", "host cell", "cell line" and "cell culture" are used interchangeably herein and all such terms should be understood to include progeny resulting from growth or culturing of a cell. "Transformation" and "transfection" are used interchangeably to refer to tlie process of introducing DNA into a cell. "Operably linlced" refers to the covalent joining of two or more nucleotide sequences, by means of enzymatic ligation or otherwise, in a configuration relative to one another such that the normal function of the sequences can be performed. For example, the nucleotide sequence encoding a presequence or secretory leader is operably hnked to a nucleotide sequence coding for a polj'peptide if it is expressed as a preprotein that participates in the secretion of the polj'peptide: a promoter or enhancer is operably linlced to a coding sequence if it affects the transcription of the sequence; a ribosome binding site is operably linked to a coding sequence if \0 'it is positioned so as to facilitate translation. Generally, "operably linked" means that the nucleotide sequences being linked are contiguous and, in the case of a secretory leader, contiguous and in reading phase. linking is accomplished by ligation at convenient restriction sites. If such sites do not exist, then synthetic oligonucleotide adaptors or linkers are used, in conjunction with standard recombinant DNA methods. The term "introduce" refers to introduction of an amino acid residue comprising an attachment group for a non-polypeptide moiety, in particular by substitution of an existing amino acid residue, or alternatively by insertion of an additional amino acid residue. The term "remove" refers to removal of an amino acid residue comprising an attachment group for a non-polypeptide moiety, in particular by substitution of the amino acid residue to be removed by another amino acid residue, or alternatively by deletion (without substitution) of the amino acid residue to be removed The term "FVn" or "FVn polypeptide" refers to a FVn molecule provided in single chain form. The term "FVDa" or "FVIIa polypeptide" refers to a FVHa molecule provided in its activated two-chain form, wherein the peptide bond between R152 and 1153 of the single-chain form has been cleaved. When the amino acid sequence of SEQ ID N0;1 is used herein to describe the amino acid sequence of FVHa it will be understood to that one of the chains '*nTVtT-*i-4Djap OTTiiT^r^ nr>i/A t-on-t/lttttp 1_1 ^O fVio /-itK^-r /^Kot-n omiTirv nr^^A TAa^Afta^a 1 ^'2 APi^ WV,/J>i^^J.XLJW/U K*.I.J.A± LAKJ UI.^.'J.^J- X VtJlWJ'WlWh? A. X W ^W] I.JLlb' \^V1.J.WX V/All*i.ii ^AXJ.AJ.Li.1^ {.AWAWl^ J. ^UX VACAWtJ J, ^f ^ T\J\J, The terms "rFVIT' and "rFVIIa" refer to FVII and FVlIa molecules produced by recombinant techniques, respectively. The terms "hFYH" and "hFVIIa" refer to wild-type human FVII and FVIIa, respectively. The term "catalytic site " is used to mean the catalytic triad consisting of S344, D242 and H193 of the FVII polypeptide. The term "active FVIIa", "active FVHa polypeptide", "active FVIIa conjugate" or "active conjugate" is used to mean a FVHa polypeptide or conjugate that possesses at least 10% of the catalytic activity of wild-type hFVIIa. Catalytic acitivity, as used herein, may suitably be determined in the assay described in the section entitled Method of measuring the catalytic acti-\nty or in the assay entitled Method of measuring low levels of catalytic activity (see the Materials and Methods section herein). Preferably, an active conjugate has at least 15%, such as at least 20%, e.g. at least 25%, more preferably at least 30%, such as at least 40%, most preferably at least 50%, e.g. at least 60% of the catalytic acitivity of wild-tj'pe hFVIIa, when tested in the assays described above. t1 Preferably, an active conjugate is able to bind to tissue factor and further activate plasma factor X and/or IX. Thus, in a preferred embodiment, the active FVIIa polypeptide or a conjugate thereof has a clotting activity of at least 25% as compared to wild ly'pe F\^a, such as a clotting activity of at least 50% as compared to wild type FVTIa, e.g. a clotting activity of at least 75% as compared to wild type FVII. Thus, the clotting activity of the active FVTIa pol}'peptide or conjugate thereof is preferably in the range of 25-200% as compared to the wild t3'pe FVHa. In particular, it is preferred that the FVHa polj'peptide or conjugate has a clotting activity in the range of 30-150% as compared to viild t}'pe FVIIa, such as a clotting activity in the range of 30-100% as compared to wild type FVIIa. The clotting activity may be determined by any method Icnown in the art as further discussed in the Materials and Methods section hereinafter. It is particularly preferred, however, that the clotting activit)' is determined in accordance with the method described in the section entitled "Method of measuring the clotting activity" (see the Materials and Method section). The term "immunogenicity" as used in connection with a given substance is intended to indicate the ability of the substance to induce a response from the immune system. The immune response may be a cell or antibody mediated response (see, e.g., Roitt: Essential Immunology (S'^ Edition, Blackwell) for further definition of immunogenicity). Normally, reduced antibody reactivity will be an indication of reduced immunogenicity. The reduced immunogenicity may be determined by use of any suitable method Icnown in the art, e.g. in vivo or in vitro. The tei-m "inactive FVHa", "inactive FVIIa polj'peptide", "inactive FVIIa conjugate" or "inactive conjugate" is used to mean a F\TIa polypeptide or conjugate that possesses less than 10% of the catalytic activity of wild-type hFVIIa. Catalytic acitivity, as used herein, may suitably be determined in the assay described in the section entitled Method of measuring the catalytic activity or in the assay entitied Method of measuring low levels of catalytic activity (see the Materials and Methods section herein). Preferably, an inactive conjugate has less than 8%, such as less than 6%, e.g. less than 5%, more preferably less than 4%, such as less than 3%, most preferably less than 2%, e.g. less than 1% of the catalytic acitivity of wild-type hFVIIa, when tested in the assays described above. Typically, an inactive conjugate has significantiy reduced in vitro or in vivo clotting activity as compared to wild-type hFVTla. The inactive F\TI or FATIa polypeptide or conjugate may be capable of competing with wild-type FVJl or FVRSL for binding tissue factor, thereby inhibiting clotting activit}'. Preferabh', the inactive YVE or F\TIa polypeptide or conjugate has less than 1% clotting acti'vity compared to wild-t}'pe hFYH. or hFXTIa. More preferably the inactive FVII orFVTEa polypeptide or conjugate has less than 0.05% clottiiig activity compared to wild type hFVH or hFVIIa. Most preferably the inactive FVH or FVHa polypeptide or conjugate has less than 0.01% clotting activity as compared to wild type hFVn or hFVTIa. In a similar way as described above, the clotting activity may be determined by any method known in the art as further discussed in the Materials and Method section hereinafter, but is preferably determined in accordance with the method described in the section entitled "Method of measuring the clotting activity". The term "functional in vivo half-life" is used in its normal meaning, i.e. the time at which 50% of the biological activity of the polypeptide or conjugate is still present in the body/target organ, or the time at which the activity of the polypeptide or conjugate is 50% of the initial value. As an alternative to determining functional in vivo half-life, "serum half-life" may be determined, i.e. the time at which 50% of the polypeptide or conjugate molecules circulate in the plasma or bloodstream prior to being cleared. Determination of serum half-life is often more simple than determining the functional in vivo half-life and the magnitude of serum half-life is usually a good indication of the magnitude of functional in vivo half-life. Alternatively terms to serum half-life include "plasma half-life", "circulating half-life", "serum clearance", "plasma clearance" and "clearance half-life". The polypeptide or conjugate is cJeared by the action of one or more of the reticuloendothelial systems (RES), kidney, spleen or liver b'" tissue factor, SBC rece-T^tor or other receT^tor mediated elimination, or bv sf^ecific or unspecific proteolysis. Normally, clearance depends on size (relative to the cutoff for glomerular filtration), charge, attached carbohydrate chains, and the presence of cellular receptors for the protein. The functionality to be retained is normally selected from procoagulant, proteolytic or receptor binding activity. The functional in vivo half-life and the serum half-life may be determined by any suitable method known in the art as further discussed in the Materials Methods section below. The term "increased" as used about the functional in vivo half-life or plasma half-life is used to indicate that the relevant half-life of the conjugate or polypeptide is statistically significantly increased relative to that of a reference molecule, such as a non-conjugated rFVUa (e.g. NovoSeven®) as determined under comparable conditions. For instance, the relevant half-life may increased by at least about 25%, such as by at least about 50%, e.g. by at least about 100%, 150%, 200%, 250%, 300%, 500% or 1000%. The term "renal clearance" is used in its normal meaning to indicate any clearance taking place by the kidneys, e.g. by glomerular filtration, tubular excretion or degradation in the 13 tubular cells. Renal clearance depends on ph}'sical characteristics of the conjugate, including size (diameter), hydrodynamic volume, symmetry, shape/rigidity, and charge. Normally, a molecular weight of about 67 IcDa is considered to be a cut-off-value for renal clearance. Renal clearance may be established by any suitable assay, e.g. an established m vivo assay. Tj'pically, renal clearance is determined by administering a labelled (e.g. radiolabelled or fluorescence labelled) polypeptide conjugate to a patient and measuring the label activity in urine collected from the patient. Reduced renal clearance is determined relative to a corresponding reference polj/peptide, e.g. the corresponding non-conjugated pol}peptide, a non-conjugated corresponding wild-tj'pe polypeptide or another conjugated polypeptide (such as a conjugated polypeptide not according to the invention), under comparable conditions. Preferably, the renal clearance rate of the conjugate is reduced by at least 50%, preferably by at least 75%, and most preferably by at least 90% compared to a relevant reference polypeptide. The ability of the conjugates of the invention to exhibit a reduced sensitivity to proteolytic degradation is of utmost importance; Compositions comprising degradation-products will typically have less specific activity as compared to compositions in which none or only a minor part of the conjugate has been degraded. Furthermore, a content of non-physiological degradation products in the composition to be administered may trigger the immune system of the patient. The term "reduced sensitivity to proteolytic degradation" is primarily intended to mean that the conjugate has reduced sensitivity to proteolytic degradation in comparison to non-conjugated wild tj'pe FVHa as determined under comparable conditions. Preferably, the proteolytic degradation is reduced by at least 10%, such as at least 25% (e.g. by 10-25%), more preferably by at least 35%, such as at least 50%, (e.g. by 10-50%, such as 25-50%) even more preferably by at least 60%, such as by at least 75% or even at least 90%. Most preferably, the proteol}tic degradation is reduced by 100%. Thus, preferably the conjugate of the invention is subjected to proteolytic degradation to a lesser extent than wild-type FVIIa, i.e. compared to non-conjugated wild tjpe FVUa the proteoljtic degradation of the conjugate of the invention is preferably reduced by 10-100%, such as by 25-100%, more preferably by 50-100%, and most preferably by 75-100%. The present inventors ha^'e de\'eloped a suitable preliminary in vitro test, which may be employed in the assessment of whether such conjugates possess reduced sensitivity to proteoljtic cleavage (reduced autoproteolysis). Thus, in a preferred embodiment of the invention the conjugate of the inN'ention has a reduced sensitivity to proteol>tic degradation (as defined above) as compared to wild t\'pe F\TIa -v^-hsn determined by the method described in the section entitled "Measurement of reduced sensitivity to proteolytic degradation", when determined by the method described in the section entitled MetJiod of Pleasuring the catalytic activity or when determined by the method described in the section entitled Method of measuring low levels of catalytic activity (see the Materials and Methods section herein). The term "parent FVn " or "parent polypeptide" is intended to indicate the molecule to be modified in accordance with the present invention. A typical parent FVII is the hFVII or hFVUa (including rFVTEa (NovoSeven®)) with the amino acid sequence shown in SEQ ID N0:1. A "variant" is a polypeptide, which differs in one or more amino acid residues from a parent polypeptide, normally in 1, 2,3,4, 5, 6,7, 8, 9,10,11,12,13,14 or 15 amino acid residues. Conjugate of the invention The conjugates of the invention are the result of a generally new strategy for developing improved FVH or FVHa molecules. More specifically, by removing and/or introducing an amino acid residue comprising an attachment group for the non-polypeptide moiety it is possible to specifically adapt the polypeptide so as to make the molecule more susceptible to conjugation to the non-polypeptide moiety of choice, to optimize the conjugation surface of the FVII or FVHa molecule and to ensure that only the attachment groups intended to be conjugated is present in the molecule) and thereby obtain a new conjugate molecule, which has or has not FVH activity and in addition one or more improved properties as compared to FVn and FVHa molecules available today. For instance, when the total number of amino acid residues comprising an attachment group for the non-polypeptide of choice is increased or decreased to an optimized level, the renal clearance of the conjugate is typically significantly reduced due to the altered shape, size and/or charge of the molecule achieved by the conjugation. In preferred embodiments of the present invention more than one amino acid residue of the FVn or FVCa polypeptide is altered, e.g. the alteration embraces removal as well as introduction of amino acid residues comprising an attachment group for the non-polypeptide^ moiety of choice. In addition to the removal and/or introduction of amino acid residues the polypeptide may comprise other substitutions or glycosylations that are not related to introduction and'or removal of amino acid residues comprising an attachment group for the IS" non-polypeptide moiety. Also, the polypeptide may be attached, e.g., to a serine proteinase inhibitor to inhibit the catal}'tic site of the polypeptide. The amino acid residue comprising an attachment group for a non-polypeptide moiety, either it be removed or introduced, is selected on the basis of the nature of the non-polypeptide moiety of choice and, in most instances, on the basis of the method in which conjugation between the polypeptide and the non-polypeptide moiety is to be achieved. For instance, when the non-polypsptide moiety is a polymer molecule such as a polyethylene glycol or polyalkylene oxide derived molecule amino acid residues comprising an attachment group may be selected from the group consisting of lysine, cysteine, aspartic acid, glutamic acid, histidine, and tjTosine, preferably cysteine and lysine, in particular lysine. 'i^Tienever an attachment group for a non-polj^eptide moiety is to be introduced into or removed from the FVH or FVHa polypeptide in accordance with the present invention, the position of the polypeptide to be modified is preferably located at the surface of the polypeptide, and more preferably occupied by an amino acid residue which has more than;25% of its side chain exposed to the solvent, preferably more than 50% of its side chain exposed to the solvent. Such positions have been identified on the basis of an analysis of a 3D structure of the human FVH or FVIIa molecule as described in the Materials and Methods section herein. Furthermore, the position is preferably selected from a part of the FVn molecule that is located . outside a -tissue factor binding site region and/or an active site region. These regions ai^e identified in the Materials and Metliods section hereinafter. It should be emphasized, however, that in certain situations, e.g. in case an inactivated conjugate is desired, it may be advantageous to perform modifications in or close to such regions. For example, it is contemplated that one or more attachment groups for the non-polypeptide moieties, such as attachment groups for in vivo N-glycosylation sites, may advantageously be inserted in the active site region or at the ridge of the active site binding cleft of the FVH molecule. The active site region and the ridge of the active site binding cleft are defined in the Materials and Methods section herein and is constituted by the following residues: 1153, Q167, V168, L169, L170, L171, Q176, L177, C178, G179, G180, T181, V188, V1S9, S190, A191, A192, H193, C194, F195, D196, K197,1198, W201, V228,1229,1230, P231, S232, T233, Y234, V235, P236, G237, T238, T239, N240, H241, D242,1243, A244, L245, L246, V28L S282, G2S3, W284, G2S5, Q2S6, T293, T324, E325, Y326, M327, F32S, D338, S339, C340, K341, G342, D343, S344, G345, G346, P347, H348, L358, T359, G360, 1361, V362, S363, W364, G365, C36S. V376, Y377, T378, R379, V380, Q382, Y383, W386, \G L387, L400 and F405 (active site region); and N173, A175, K199, N200, N203, D289, R290, G291, A292, P321 and T370 (the ridge of the active site binding cleft). In order to determine an optimal distribution of attachment groups, the distance between amino acid residues located at the surface of the FVn or FVHa molecule is calculated on the basis of a 3D structure of the polypeptide. More specifically, the distance between the CB's of the amino acid residues comprising such attachment groups, or the distance between the functional group (NZ for lysine, CG for aspartic acid, CD for glutamic acid, SO for cysteine) of one and the CB of another amino acid residue comprising an attachment group are determined. In case of glycine, CA is used instead of CB. In the FVH or FVTIa polypeptide part of a conjugate of the invention, any of said distances is preferably more than 8 A, in particular more than lOA in order to avoid or reduce heterogeneous conjugation. In case of removal of an attachment group, the relevant amino acid residue comprising such group and occupying a position as defined above is preferably substituted with a different amino acid residue that does not comprise an attachment group for the non-polypeptide moiety in question. Normally, the amino acid residue to be removed is one to which conjugation is disadvantageous, e.g. an amino acid residue located at or near a functional site of the polypeptide (since conjugation at such a site may result in inactivation or reduced FVTI or context the term "functional site" is intended to indicate one or more amino acid residues which is/are essential for or otherwise involved in the function or performance of FVU or FVIIa. Such amino acid residues are a part of the functional site. The functional site may be determined by methods known in the art and is preferably identified by analysis of a structure of the FVlIa-iissue factor complex (See Baimer et al.. Nature 1996; 380:41-46). In case of introduction of an attachment group, an amino acid residue comprising such group is introduced into the position, preferably by substitution of the amino acid residue occupying such position. The exact number of attachment g"oups present and available for conjugation in the , E^Vn or FVIIa polypeptide is dependent on the effect desired to be achieved by the conjugation. The effect to be obtained is, e.g., dependent on the nature and degree of conjugation (e.g. the identity of the non-polypeptide moiet}', the number of non-polypeptide moieties desirable or possible to conjugate to the polypeptide, where they should be conjugated or where conjugation should be avoided, etc.). \1 Functional in vivo half-life is i.a. dependent on the molecular weight of the conjtigate, and the number of attaciiment groups needed for providing increased half-life thus depends on the molecular weight of the non-polypeptide moiety in question. In one embodiment, the conjugate of the invention has a molecular weight of at least 67 IcDa, in particular at least 70 IdDa, e.g., as measured by SDS-PAGE according to Laemmli, U.K., Nature Vol 227 (1970), p680-85. FVn has a molecular weight of about 53 kDa, and therefore additional 10-20kDa is required to obtain the desired effect. This may, e.g., be provided by conjugating 2-4 lOkDa PEG molecules or as otherwise described herein. In order to avoid too much disruption of the strucmre and function of the parent molecule the polypeptide part of the conjugate will typically have an amino acid sequence having more than 90% identity with SEQ ID N0:1, preferably more than 95%, such as more than 96%. In particular, the polj'peptide part of the conjugate will typically have an amino acid sequence having more than 97% identity with SEQ ID N0:1, such as more than 98%, more than 99%, more than 99.25%, more than 99.25% or more than 99.5%. Amino acid sequence homology/identity is conveniently determined from aligned sequences, using e.g. the ClustalW program, version 1.8, June 1999, using default parameters (Thompson et al., 1994, ClustalW: Improving the sensitivity of progressive multiple sequence alignment tlirough sequence weighting, position-specific gap penalties and weight matrix choice, Nucleic Acids Research, 22: 4673-4680) or from the PFAM families database version 4.0 rhttp://pfam.wustl.edu/) (Nucleic Acids Res. 1999 Jan 1; 27(l):260-2) by use of GENEDOC version 2.5 (Nicholas, K.B., Nicholas H.B. Jr., and Deerfield, D.W. E. 1997 GeneDoc: Analysis and Visualization of Genetic Variation, EMBNEW.NEWS 4:14; Nicholas, K.B. and Nicholas H.B. Jr. 1997 GeneDoc: Analysis and Visualization of Genetic Variation). Stated differently, the total number of amino acid residues to be altered in accordance with the present invention (as compared to the amino acid sequence shown in SEQ DD N0:1) will typically not exceed 15. Preferably, the FVJl or FVUa polj'peptide part of the conjugate of the invention or the polypeptide of the invention comprises an amino acid sequence differing in 1-15 amino acid residues from the amino acid sequence shown in SEQ ID N0:1, typically in 1-10 or in 2-10 amino acid residues, e.g. in 1-8 or in 2-8 aniino acid residues, such as in 3-7 or in 4-6 amino acid residues from the amino acid sequence shown in SEQ ID N0:1. Thus, normally the pol>'peptide part of the conjugate or the poh'peptide of the invention comprises an amino acid sequence which differs from the amino acid sequence shown in SEQ ID N0:1 in at the most 15 aniino acid residues (such as 15 amino acid residues), in at the most 14 amino acid residues (such as 14 amino acid residues), in at the most 13 amino acid residues (e.g. 13 amino acid residues), in at the most 12 amino acid residues (such as 12 amino acid residues), in at the most 11 amino acid residues (such as 11 amino acid residues), in at the most 10 amino acid residues (e.g. 10 amino acid residues), in at the most 9 amino acid residues (such as 9 amino acid residues), in at the most 8 amino acid residues (such as 8 amino acid residues), in at the most 7 amino acid residues (such as 7 amino acid residues), in at the most 6 amino acid residues (such as 6 amino acid residues), in at the most 5 amino acid residues (such as 5 amino acid residues), in at the most 4 amino acid residues (such as 4 amino acid residues), in at the most 3 amino acid residues (such as 3 amino acid residues) or in at the most 2 amino acid residues (such as 2 amino acid residues). Analogously, the conjugate of the invention typically contains, e.g., 1-15 non-polypeptide moieties, typically 1-10 non-polypeptide moieties or 2-10 non-polypeptide moieties, such as 1-8 or 2-8 non-polypeptide moieties, e.g. 1-6,1-4, 3-7 or 4-6 non-polypeptide moieties. Preferably, the conjugate of the invention has one or more of the following improved properties: Increased functional in vivo half-life, increased plasma half-life, reduced renal clearance and reduced sensitivity to proteolytic degradation as compared to rFVIIa (e.g.NovoSeven®). It is known from i.a. WO 88/10295 that proteolytic degradation of FVH/FVlIa ^xxLLiuxxiy Lttivuo jjicw^v^ at vaiiuu.o piL/tt>ui^tiv^ oiLt/o ui tii^ xxiui&wuio, iiouioi^ at uio pu&iuuiia x\.^^} K38,142, Y44, K143, R290, R315, K341, R392, R396 and R402 (or rather between the positions K32-D33, K38-L39,142-S43, Y44-S45, K143-R144, R290-G291, R315-K316, K341- G342, R392-S393, R396-P397 and R402-A403). Thus, one preferred strategy for increasing e.g. the functional in vivo half-life, the increased plasma half-life or for reducing the sensitivity to proteolytic degradation is to modify the parent polypeptide at and/or around one or more of these proteolytic degradation sites, e.g. by introducing non-polypeptide moieties at and/or around these sites. Thus, in a preferred embodiment of the invention an attachment group has been introduced in one or more of the above-mentioned identified positions or at position -4, -3, -2, -1,1,2,3,4, preferably position -2, -1,1,2, such as position -1,1, relative to the position directly involved in proteolytic degradation. L More specifically, it is preferred that a non-naturally occurring in vivo glycosylation site, in particular a non-naturally occurring in vivo N-glycosylation site (see further below) has been introduced in positions selected from the group consisitng of 28-48,139-147,286-294, 311-319,338-345, and 388-406. In particular, it is preferred that the in vivo glycosylation site, R such as an in vivo N-glycosylation site (see further below) has been introduced in positions selected from the group consisiting of 30-34, 36-46,141-144,288-292, 313-317, 341-343, 390-398 and 400-404. More preferably, the in vivo glycosylation site, such as an in vivo N-glycosjdation site (see further below) has been introduced in positions selected from the group consisiting of 31-33 (e.g. positions 31, 32 or 33), 37-39 (e.g. positions 37,38 or 39), 41-46 (e.g. positions 41,42, 43,44,45 or 46), 142-144 (e.g. positions 142,143 or 144), 289-291 (e.g. positions 289, 290 or 291), 314-316, (e.g. positions 314, 315 or 316), 341-342 (e.g. positions 341,342 or 343), 391-393 (e.g. positions 391, 392 or 393), 395-397 (e.g. positions 395, 396 or 397) and 401-403 (e.g. positions 401, 402 or 403). The introduction is preferably performed by susbstitution. Conjugate of the invention, wherein the non-polypeptide moietyAs a molecule tliat has lysine as an attachment group In an interesting embodiment of the invention the non-polypeptide moiety has-lysine as an attachment group, i.e., the conjugate of the invention is one which comprises at least one non-polypeptide moiety covalently attached to a polypeptide, wherein the amino acid sequence of the polypeptide differs from that of wild-type FVII or FVlIa shown in SEQ ID NO: 1 in that y at least one lysine residue has been introduced or removed. FVn/FVIIa contains 17 lysine residues. Three lysine residues (K18, K62 and K85) are located in the tissue factor binding domain and two lysine residues (K197 and K341) are located in the active site region. Due to the relative high amount of lysine residues in the parent polypeptide it is envisaged that at least one lysine residue should preferably be removed, in particular by substitution of a lysine residue with a non-lysine residue, in order to avoid excessive conjugation to non-polypeptide moieties. Thus, in one embodiment the amino acid sequence of the FVII or FVIIa pol}'peptide part of the conjugate differs from that shown in SEQ ID NO-.l in that at least one lysine residue, such as 1-15 lysine residues, in particular 1-10,1-6 or 2-4 lysine residues, has been removed, preferably by substitution. For example, the lysine residue(s) to be removed, preferably by substitution, is selected from the group consisting of K18, K32, K38, K62, K85, K109, K137, K143, K148, K157, K16L K197, K199, K316, K337, K34L K389 and combinations thereof. In particular, it is preferred to remove one or more lysine residues, which constitute part of tissue factor binding site and/or the active site region, i.e. the residues K18, K62, K85, K197, K341 or ^C combinations thereof. The lysine residue may be substituted with any other amino acid residue, but is preferably substituted with R, Q, N or H, more preferably R In another embodiment the amino acid sequence of the FVn or FVHa polypeptide part of the conjugate differs from that shown in SEQ ID N0:1 in that at least one lysine residue, such as 1-15 lysine residues, in particular 1-10,1-6 or 2-4 lysine residues, has been introduced, preferably by substitution. It will be understood that it is particularly preferred that at least one of the lysines residues, which are introduced at a predetermined site in the parent molecule, is combined with at least one of the above-mentioned removals of lysine residues. Thus, in a preferred embodiment of the invention, the amino acid sequence of the FVH or FVUa polypeptide part of the conjugate differs from that shoviii in SEQ ID N0:1 in that at least one lysine residue has been removed and at least one lysine residue has been introduced. Examples of positions, wherein lysine residues may be introduced include, but is not limited to, positions at or in the vicinity of the proteolytic degradation sites described above. Thus, in a preferred embodiment the substitution of a non-lysine residue vvdth a lysine residue is selected from tiie group consisting of of I42K, Y44K, L288K, D289K, R290K, G291K, A292K, T293K, Q313K, S314K, R315K, V317K, L390K, M391K, R392K, S393K, E394K, P395K, R396K, P397K. G398K, V399K, L400K, L401K, R402K, A403K, P404K, F405K and combinations thereof, in particular selected from the group consisting of R290K, R315K, R392K, R396K, R402K and combinations thereof. While the non-polypeptide moiety of the conjugate according to this aspect of the invention may be any molecule which, when using the given conjugation method has lysine as an attachment group it is preferred that the non-polypeptide moiety is a polymer molecule. The polymer molecule may be any of the molecules mentioned in the section entitled "Conjugation to a polymer molecule", but is preferably selected from the group consisting of linear or branched polyethylene glycol or another polyalkylene oxide. Examples of preferred polymer molecules are, e.g., SS-PEG, NPC-PEG, aldehyde-PEG, mPEG-SPA, mPEG-SCM, mPEG-BTC from Shearwater Polymers, Inc, SC-PEG from Enzon, Inc., tresylated mPEG as described in US 5,880,255, or oxycarbonyl-oxy-N-dicarboxyimide-PEG (US 5,122,614). Normally, for conjugation to a lysine residue the non-pol}'peptide moiety has a molecular weight of from about 5 to about 20 kDa, such as from about 5 to about 10 KDa, e.g. about 5 kDa or about 10 kDa It will be understood that any of the amino acid changes, in particular substitutions, specified in this section can be combined with any of the amino acid changes, preferably ai substitutions specified in the other sections herein disclosing specific amino acid modifications, including introduction and/or removal of glycosylation sites. Conjugate of the invention wherein the non-polypeptide moiety is a molecule that has cysteine as an attachment group In a further interesting embodiment of the invention the non-polypeptide moiety has cysteine as an attachment group, i.e., the conjugate of the invention is one wliich comprises at least one non-pol5'peptide moiety covalently attached to a polypeptide, wherein the amino acid sequence of the pol}'peptide differs from that of \^ild-type FVH or FVUa shown in SEQ ID NO: 1 in that at least one cj'steine residue has been introduced or removed. FSm/FVIIa contains 24 cysteine residues and disulfide bridges are established between the following cysteine residues; C17 and C22, C50 and C61, C55 and C70, C72 and C81, C91 and C102, C98 and C112, C114 and C127, C135 and C262, C159 and C164, C178 and C194, C310 and C329, and between C340 and C368. ,, r In a further interesting embodiment, the amino acid sequence of the FVn or FVIIa polypeptide differs from that shown in SEQ E) N0:1 in that at least one cysteine residue, such as 1-15 cysteine residues, in particular 1-10, 1-6 or 2-4 cysteine residues, has been introduced, preferably by substitution. Examples of positions, wherein cysteine residues may be introduced include, but is not limited to, positions at or in the vicinity of the proteol>'tic degradation sites described above. Thus, in an interesting embodiment of the invention the^lysine residue(s)l:o'be ' introduced, preferably by substitution, is selected from the group consisting of BOC, K32C, D33C, A34C, T37C, K38C, W41C, Y44C, S45C, D46C, L141C, E142C, KI43C, R144C, L288C, D289C, R290C, G291C, A292C, S314C, R315C, K316C, V317C, L390C, M391C, R392C, S393C, E394C, P395C, R396C, P397C, G398C, V399C, L401C, R402C, A403C, P404C and combinations thereof, in particular selected from the group consisting of K32C, Y44C, K143C, R290C, R315C, K34IC, R392C, R396C, R402C and combinations thereof. In a further interesting embodiment of the invention the cysteine residue(s) is/are introduced into a position that in wildtj'pe hF\m is occupied by a threonine or serine residue having at least 25% of its side chain exposed to the surface. For instance, in the FVH or FXTIa polypeptide a cysteine residue is introduced, preferably by substitution, into at least one position selected from the group consisting of S12, S23, S43, S45, S52, S53, S60, S67, T83, S103, T106, T108, Sill, S119, S126, T128, T130, S147, T185, S214, S222, S232, T233, T238, T239, T255, T267, T293, T307, S320, T324, S333, S336, T370 and S393. Even more ^ • preferably the cysteine residue is introduced into at least one position of hFVU containing an S residue, the position being selected from the group consisting of S12, S23, S43, S45, S52, S53, S60, S67, S103, Sill, S119, S126, S147, S214, S222, S232, S320, S333, S336 and S393. In a further embodiment the cysteine residue(s) is/are introduced into a position that in wildtype hFVn is occupied by a threonine or serine residue having at least 50% of its side chain exposed to the surface. For instance, in the FVn or FVHa polypeptide a cysteine residue is introduced, preferably by substitution, into at least one position selected from the group consisting of S23, S43, S52, S53, S60, S67, T106, T108, Sill, SI 19, S147, S214, T238, T267 and T293, even more preferably a position selected from the group consisting of S23, S43, S52, S53, S60, S67, Sill, S119, S147 and S214. In a still further embodiment a cysteine residue is introduced into at least one position selected from any of the above-mentioned positions, which is not located in an active site region. Preferably, the position is one occupied by a T or an S residue. As an example, the FVII polypeptide comprises a cysteine residue introduced into at least one position selected from the group consisting of S12, S23, S43, S45, S52, S53, S60, S67, T83, S103, T106, T108, Sill, S119, S126, T128, T130, S147, T185, S214, S222, T255, T267, T307, S320, S333, S336, T370 and S393 (having more than 25% of its side chain exposed to the surface), in particular selected from the group consisting of S12, S23, S43, S45, S52, S53, S60, S67, S103, Sill, S119, S126, S147, S214, S2.2.2, S320, S333; S33fi and S393 (occupied by S residue), and more preferably from the group consisting of S23, S43, S52, S53, S60, S67, T106, T108, Sill, S119, S147, S214 and T267 (having more than 50% of its side chain exposed to the surface), in particular from the group consisting of S23, S43, S52, S53, S60, S67, Sill, S119, S147 and S214 (occupied by an S residue). In an even further embodiment a cysteine residue is introduced into at least one position selected from any of the above lists, which is not located in a tissue factor binding site region. Preferably, the position is one occupied by a T or an S residue. As an example, the FVII polypeptide comprises a cysteine residue introduced into at least one position selected from the group consisting of S12, S23, S45, S52, S53, S67, T83. S103, T106, T108, Sill, S119, S126, T128, T130, S147, T185, S214, S222, S232, T233, T238, T239, T255, T267, T293, S320, T324, S333, S336, T370 and S393 (having more than 25% of its side chain exposed to the surface), in particular selected from the group consisting of S12, S23, S45, S52, S53, S67, S103, SlU, S119, S126, S147, S214, S222, S232, S320, S333, S336 and S393 (occupiedby S residue), and more preferably from the group consisting of S23, S52, S53, S67, T106, T108, Sill, S119, S147, S214, T238, T267 and T293 (having more than 50% of its side chain exposed to the surface), in particular from the group consisting of S23, S52, S53, S67, Sill, SI 19, S147 and S214 (occupied by an S residue). In a still further embodiment a cysteine residue is introduced into at least one position selected from any of the above lists, which is neither located in a tissue factor binding site region nor in an active site region. Preferably, the position is one occupied by a T or an S residue. As an example, the FVn polypeptide comprises a cysteine residue introduced into at least one position selected from the group consisting of S12, S23, S45, S52, S53, S67, TS3, S103, T106, TIGS, Sill, S119, S126, T128, T130, S147, T1S5, S214,'S222, T255, T267, S32D, S333, S336, T370 and S393 (having more than 25% of its side chain exposed to the surface), in particular selected from the group consisting of S12, S23, S45, S52, S53, S67, S103, Sill, Sn9, S126, S147, S214, S222, S320, S333, S336 and S393 (occupied by S residue), and more preferably from the group consisting of S23, S52, S53, S67, T106, TIOS, Sill, S119, S147, S214 and T267 (having more than 50% of its side chain exposed to the surface), in particular from the group consisting of S23, S52, S53, S67, Sill, S119, S147 and S214 (occupied by an S residue). "VsTiile the non-polypeptide moiety of the conjugate according to this aspect of the invention may be any molecule which, when using the given conjugation method has cysteine as an attachment group it is preferred that the non-polypeptide moiety is a polymer molecule. The polymer molecule may be any of the molecules mentioned in the section entitled "Conjugation to a polymer molecule", but is preferably selected from the group consisting of linear or branched polyethylene glycol or another polyalkylene oxide. In a particular interesting embodiment the polymer molecule is PEG, such as VS-PEG. The conjugation between the polypeptide and the polymer may be achieved in any suitable manner, e.g. as described in the section entitled "Conjugation to a polj'mer molecule", e.g. in using a one step method or in the stepwise manner referred to in said section. When the FVIl or FVIIa polypeptide comprises only one conjugatable cysteine residue, this is preferably conjugated to a non-polypeptide moiety with a molecular weight of from about 5 MDa to about 20 IcDa, e.g. from about 10 kDa to about 20 IvDa, such as a molecular weight of about 5 kDa, about 10 kDa, about 12 kDa, about 15 IdDa or about 20 IdDa, either directly conjugated or indirectly through a low molecular weight polymer (as disclosed in WO 99/55377). When the conjugate comprises tv«'o or more conjugatable cysteine residue, normally each of the non-polypeptide moieties has a molecular weight of from about 5 to about 10 kDa, such as about 5 IcDa or about 10 IcDa. It mil be understood that any of ihe amino acid changes, in particular substimtions, specified in this section can be combined uiih an)' of the amino acid changes, preferably 'substitutions specified in the other sections herein disclosing specific amino acid modifications, including introduction and/or removal of glycosylation sites. Conjugate of the invention wherein the non-polypeptide moiety is a molecule that has aspartic acid or glutamic acid as an attachnent group. In a still further interesting embodiment of the invention the non-polypeptide moiety has aspartic acid or glutamic acid as an attachment group, i.e., the conjugate of the invention is one which comprises at least one non-polypeptide moiety covalently attached to a polypeptide, wherein the amino acid sequence of the polypeptide differs from that of wild-type FS^ or FVHa shown in SEQ E) NO: 1 in that at least one aspartic acid residue and/or at least one glutamic acid residue has been introduced or removed. In a further interesting embodiment, the amino acid sequence of the FVH or FVJL& polypeptide differs from that shown in SEQ ID N0:1 in that at least one aspartic acid residue and/or glutamic acid residue, such as 1-15 aspartic acid residues and/or glutamic acid residues, in particular 1-10,1-6 or 2-4 aspartic acid residues and/or glutamic acid residues, has been introduced, preferably by substitution. Examples of positions, wherein aspartic acid residues or glutamic acid residues may be introduced include, but is not limited to, positions at or in the vicinity of the proteolytic degradation sites described above. Thus, in an interesting embodiment of the invention the aspartic acid residue and/or the glutamic acid residue to be introduced, preferably by substitution, is selected from the group consisting of I30D/E, K32D/E, A34D/E, T37D/E, K38D/E, W41D/E, Y44D/E, S45D/E, D46C, L141D/E, E142D/E, K143D/E, R144D/E, L288D/E, R290D/E, G291D/E, A292D/E, Q313D/E, S314D/E, R315D/E, K316D/E, V317D/E, L390D/E, M391D/E, R392D/E, S393D/E, P395D/E, R396D/E, P397D/E, G398D/E, V399D/E, L401D/E, R402D/E, A403D/E, P404D/E, and combinations thereof, in particular selected from the group consisting of K32D/E, Y44D/E, K143D/E, R290D/E, R315D/E, K341D/E, R392D/E, R396D/E. R402D/E and combinations thereof. In addition to the above listed substitution(s), the polypeptide of the conjugate according to the above embodiment may comprise removal, preferably by substitution, of at least one of the aspartic acid residue and/or at least one glutamic acid residue. Due to the relative high amount of lysine residues in the parent polypeptide it is envisaged that at least one aspartic acid or glutamic acid residue should preferably be removed, 'IS' ' in particular by substitution, in order to avoid excessive conjugation to non-polypeptide moieties. Thus, in one embodiment tlie amino acid sequence of the FVII or FVHa polypeptide part of the conjugate differs from that shown in SEQ ID N0;1 in that at least one aspartic acid or glutamic acid residue, such as 1-15 aspartic acid or glutamic acid residues, in particular 1-10, 1-6 or 2-4 aspartic acid or glutamic acid residues, has been removed, preferably by substitution. For example, the aspartic acid and glutamic acid residue(s) to be removed, preferably by substitution, is selected from the group consisting of D33, D46, D4S, E77, E82, D86, D87, E94, E99, D104, E116, D123, E132, E142, E163, D196, E210, D212, E215, D217, D219, E220, D256, E265, E270, D289, E296, D309, D319, E325, D334, D338, D343, E385, E394 and combinations thereof. It will be understood that it is particularly preferred that at least one of the aspartic acid or glutamic acid residues, which are introduced at a predetermined site in the parent molecule, is combined with at least one of the above-mentioned removals of aspartic acid or glutamic acid residues. Thus, in a prefen'ed embodiment of the invention, the amino acid sequence of the FVn or FVTIa polypeptide part of the conjugate differs from that shown in SEQ ID N0:1 in that at least one aspartic acid or glutamic acid residue has been removed, preferably by substitution, and at least one aspartic acid or glutamic acid residue has been introduced, preferably by substitution. While the non-polypeptide moiety of the conjugate according to this aspect of the invention, which has an aspartic acid group or a glutamic acid group as an attachment group, can be any non-polypeptide moiety with such property, it is presently preferred that the non-pol3'peptide moiety is a polymer molecule or an organic derivatizing agent, in particular a polymer molecule, and the conjugate is prepared, e.g., as described by Salcane and Pardridge, Pharmaceutical Research, Vol. 14, No. 8, 1997, pp 1085-1091. It will be understood that any of the amino acid changes, in particular substitutions, specified in this section can be combined with any of the amino acid changes, in particular substitutions specified in the other sections herein disclosing specific amino acid changes, including introduction and/or removal of glycosylation sites. Conjugate of the inventi.07i wherein the non-polypeptide moiet)' is a sugar moiety In a further interesting embodiment of the invention, an attachment group for a sugar moiety, such as a glycos}'lation site, in particular an in vivo glycosylation site, has been inserted and/or remoA'ed. rrereraoiy, me conjugate or tne mvennon is one wmcn comprises ai jeasr one sugar 3ioiety covalently attached to a polypeptide, wherein the amino acid sequence of the polypeptide differs from that of wild-type FVn or FVIIa shown in SEQ ID N0:1 in that at least Dne non-naturally occurring glycosylation site has been introduced and/or at least one naturally Dccurring glycosylation site has been removed. In particular, a non-naturally occurring ^ycosylation site has been introduced, or a non-naturally occurring glycosylation site has been ntroduced in combination with the removal of a natural occurring glycosylation site. The ntroduced glycosylation site may be an 0-glycosylation site or an N-glycosyiation site, ^'referably the glj'cosylation site is an in vivo 0-glycosylation site or an in vivo N-glycosylation iite, in particular an in vivo N-glycosylation site. When used in the present context, the term "naturally occurring glycosylation site" covers the glycosylation sites at postions N145, N322, S52 and S60. In a similar way, the term 'naturally occurring in vivo O-glycosylation site" includes the positions S52 and S60, whereas he term "naturally occurring in vivo N-glycosylation site" includes positions N145 and N322. Typically, the amino acid sequence of the FVn or FVHa polypeptide differs from that ;hown in SEQ ID N0:1 in that at least one non-naturally occurring glycosylation site (e.g. at east one non-naturally occurring in vivo N-glycosylation site), such as 1-15 non-naturally )ccurring glycosylation sites (e.g. 1-15 non-naturally occurring in vivo N-glycosylation sites), n particular 1-lOj 1-6 or 2-4 non-naturally occurring glycosylation sites (e.g. 1-10,1-6 or 2-4 \on-naturally occurring in vivo N-glycosylation sites), has been introduced, preferably by '.ubstitution. It will be understood that in order to prepare a conjugate, wherein the polypeptide of he conjugate comprises one or more glycosylation sites, the polypeptide must be expressed in a lost cell capable of attaching sugar (oligosaccharide) moieties at the glycosylation site(s) or iltematively subjected to in vitro glycosylation. Examples of glycosylating host cells are given ,n the section further below entitied "Coupling to a sugar moiety". In an interesting embodiment of this aspect, an in vivo glycosylation site is introduced .nto a position of the parent FVU or FVIIa molecule occupied by an amino acid residue exposed :o the surface of the molecule, preferably with more than 25% of the side chain exposed to the jolvent, in particular more than 50% exposed to the solvent (these positions are identified in the Methods section herein). The N-glycosylation site is then introduced in such a way that the N-residue of said site is located in said position. Analogously, an O-glycosylation site is LUtroduced so that the S or T residue making up such site is located in said position. Examples ^*r of such positions include iL52ii/i, 14Z^/1, i ^4^^/1, X^X^DCH i, ss.^:^^^/ ±, ±\^^^^, T, K341S/T, R392S/T, R396S/T, R402S/T and combinations thereof. With respect to N-glycosylation, the in vivo glycosylation site is introduced into a position wherein only one mutation is required to create the site (i.e. where any other amino acid residues required for creating a functional glycosylation site is already present in the molecule). in other \^'ords, the conjugate according to the invention is preferably a conjugate, wherein the amino acid sequence of the polypeptide differs from SEQ ID N0:1 in that at least one naturally occurring N-X'-X sequence is substituted with a N-X'-S or N-X'-T sequence, wherein X' is any amino acid except P, and X is any amino acid except for S and T. In a similar way, the conjugate according to the invention is preferably a conjugate, wherein the amino acid sequence of the polypeptide differs from SEQ ID N0:1 in that at least one naturally occuning X-X'-S or X-X'-T sequence naturally present in SEQ ID N0:1 is substituted with a N-X'-S or a N-X'-T sequence, wherein X' is any amino acid except P, and X is any amino acid except for N. Specific examples of such substitutions creating an in vivo N-glycosylation site include a substitution selected from the group consisting of F4S/T, PI ON, Q21N, W41N, S43N, A51N, G58N, L65N, G59S/T, E82S/T, N95S/T, G97S/T, YIOIN, D104N, T106N, K109N, G117N, G124N, S126N, T128N, A175S/T, G179N, I186S/T, V188N, R202S/T, I205S/T, D212N, E220N, I230N, P23 IN, P236N, G237N, V253N, E265N, T267N, E270N, R277N, L280N, ; G291N, P303S/T, L305N, Q312N, G318N, G331N, D334N, K337N, G342N, H348N, R353N, 'i Y357N, D61N, V376N, R379N, M391N, and combinations thereof. Preferably, the substimtion-' is selected from the group consisting of F4S/T, PION, Q21N, W41N, A51N, G58N, G59S/T, N95S/T, G97S/T, YIOIN, D104N, T106N, K109N, G117N,^G124N, S126N, T128N, A175S/T, I186S/T, V188N, R202S/T, I205S/T, D212N, E220N, V253N, E265N,-T267N, E270N, L280N, G291N, P303S/T, G318N, G331N, D334N, K337N, R353N, Y357N, M391N, and combinations thereof. Alternatively, the in vivo glycosylation site is introduced in a position occupied by a lysine residue or an arginine residue, preferably occupied by an lysine residue, in particular so that the N-residue of the N-glycosylation site or the S or T residue of an O-glycosylation site substitutes the lysine residue. Stated differentiy, tlie conjugate according to the invention is preferably a conjugate, wherein the amino acid sequence of the polypeptide differs from SEQ ID NO: 1 in that at least one K-X'-X sequence or R-X'-X sequence, preferably a K-X'-X sequence, naturally present in ^S SEQ E) N0:1 is substituted with a N-X'-S or a N-X'-T sequence, wherein X' is any amino acid except P, and X is any amino acid except for S and T. Specific examples of such substitutions creating an in vivo N-glycosylation site include a substitution selected from the group consisting of K32N+A34S/T, K38N+F40S/T, K62N+Q64S/T, K85N+D87S/T, K137N+P139S/r, K143N+N145S/T, K148N+Q149S/T, K161N+E163S/T, K197N+K199S/T, K199N+W201S/T, K316N+G318S/T, K337N, K341N+D343S/T, K389N+M391S/T and combinations thereof. In a further interesting embodiment of the glycosylation aspect described above, a glycosylation site, in particular an N-glycosylation site, may be introduced in positions at or in the vicinity of the proteolytic degradation sites described above (see the section entitled "Conjugate of the invention"). Thus, specific examples of such substitutions creating an in vv\>o N-glycosylation site include a substitution selected from the group consisting of K32N+A34S/T, F31N+D33Sn:, B0N+K32S/r, A34N+R36S/T, K38N+F40S/T, T37N+L39S/T, R36N+K38S/T, L39N+W41S/T, F40N+I42S/T, W41N, I42N+Y44S/T, S43N, Y44N+D46S/r, S45N+G47S/r, D46N+D4SS/T, G47N+Q49S/T, Ki43N+N145S/T, E142N+R144S/T, L141N+K143S/T, I140N+E142S/T, R144N+A146S/T, A146N+K148S/T, S147N-hP149S/T, R290N+A292S/T, D289N+G291S/T, L288N+R290S/T, L287N+D289S/T, G291N, A292N+A294S/T, T293N+L295S/T, R315N-hy317S/T, S314N+K316S/T, 0313N-i-R315S/T, 0312N, K316N+G318S/T, V317N+D319S/T, G318N, K341N+D343S/r, S339N+K341S/r, G342N, D343N+G345S/T, R392N+E394S/T, M391N, L390N+R392S/T, K389N+M391S/T, S393N+P395S/T, E394N+R396S/T, P395N+P397S/T, R396N+G398S/T, P397N+V399S/T, G398N-F-L400S/T, V399N+L401S/T, L400N+R402S/T, L401N+A403S/T, R402N+P404S/T, A403N+F405S/T, P404N+P406S/T and combinations thereof, such as K143N+N145S/T+ R315N+V317S/T. Preferably, the substitution is selected from the group consisting of K32N+A34S/T, K38N+F40S/T, Y44N+D46S/T, K143N+N145S/T, R290N+A292S/T, R315N+V317S/T, K341N+D343S/T, R392N+E394S/T, R396N+G398S/T, R402N+P404S/T and combinations thereof, such as K143N+N145S/T+ R315N+V317S/T. More preferably, the substitution is selected from the group consisting of K32N-f-A34T, K38N+F40T, Y44N+D46T, K143N+N145T, R290N+A292T, R315N+V317T, 341N+D343T, R392N+E394T, R396N+G398T, R402N-1-P404T and combinations thereof, in particular K143N+N145T-)-R315N+V317T. In a still furter interesting embodiment of the invention it is preferred that the in vivo glycosylation site is introduced in a position which does neither form part of the tissue factor binding nor form part of the active site region and the ridge of the acti^'e site binding cleft as defined herein. It is envisaged that such glycosylation variants will primarily belong to the class of active conjugates as defined hereinbefore. Thus, specific examples of substitutions creating such an in vivo N-glycosylation site include substitutions selcted from the group consisting of K32N+A34S/T, B0N+K32S/T, A34NH-R36S/T, K38N+F40S/T, T37N+L39S/T, W41N, Y44N+D46S/T, S45N+G47S/T, D46N+D48S/r, G47N+Q49S/T, KM3N+H145S/T, E142N+R144S/T, L141N+K143S/T, I140N+E142S/T, R144N+A146S/T, A146N+K148S/T, S147N+P149S/T, L288N+R290S/T, L287N+D2S9S/T, R315N+V317S/T, S314N+K315S/T, K316N+G31SS/T, V317N+D319S/T, G318N, R392N+E394S/T, M391N, L390N+R392S/T, K389N+M391S/T, S393N+P395S/T, E394N+R396S/T, P395N+P397S/T, R396N+G398S/r, P397N+V399S/T, G398N+L400S/T, V399N+L401S/T, L401N+A403S/T, R402N+P404S/T, A403N+F405S/T, P404N+P406S/T and combinations thereof, such as K143N+N145S/T+R315N+V317S/T. Preferably, the substitution is selcted from the group consisting of K32N+A34S/T, K3SN+F40S/T, Y44N+D46S/T, K1_4^N+N14^5S/T, R315N+Y317S/T, R392N+E394S/T, R396N+G398S/T, R402N+P404S/T and combinations thereof, such as K143N+N145S/T+ R315N+V317S/T. More preferably, the substitution is selected from the group consisting of K32N+A34T, K38N+F40T, Y44N+D46T, K1,43N+N145T, R3JiN+y317T, R392N+E394T, R396N+G398T, R402N+P404T and combinations thereof, in particular K143N+N145T+ R315N+V317T. In an even furter interesting embodiment of the invention it is preferred that the in vivo glycosylation site is introduced in a position which does not form part of the tissue factor but wliich forms part of the active site region and the ridge of the active site binding cleft as defined herein. It is envisaged that such glj'cosylation variants will primarily belong to the class of inactive conjugates as defined hereinbefore. Thus, specific examples of substitutions creating such an in vivo N-glycosylation site include substitutions selcted from the group consisting of I153N-fG155S/T, Q167N+L169S/T, V168N+L170S/T, L169N+L171S/T, L170N+V172S/T, L171N+N173S/T, A175S/T, A175N+L177S/T, L177N+G179S/T, G179N, G180N+L182S/T, T181N+I1S3S/T, V188N, Y189N+A191S/T, S190N+A192S/T, A191N+H193S/T, H193N+F195S/T, F195N+K197S/T, D196N+I198S/T, K197N+K199S7T, I198N+N200S/T, K199N+W201S/T, W201N+N203S/T, R202S/T, n05S/T, V228N+I230S/T, I229N+P231S/T, I230N, P231N, S232N+Y234S/T, T233N-I-V235S/T, Y234N+P236S/T, V235N-rG237S/T, P236N, G237N, T238N+N240S/T, T239N+H241S/T, H241N^I243S/T, D242S/T, I243N+L245S/T, A244N4-L246S/T, L245N+R247S/T, L246N+L246S/T, Y281N+G2S3S/T, S282N+W284S/T, G2S3N+G285S/T, 3D sV284N+Q286SyT, G285N+L287S/T, Q286N+L288S/T, D289N+G291S/T, R290N+A292S/r, 3291N, A292N+A294S/T, T293N+L295S/T, P321N+I323S/T, T324N-FY326S/T, 5325N+M327S/T, Y326N+F327S/T, F328N+A330S/T, S339N+K341S/T, K341N+D343S/r, 3342N+S344S/T, D343N+G345S/T, S344NH-G346S/T, G345N4-P347S/T, P347N+A349S/T, :-B48N, L358N+G360S/T, T359N+D61S/T, G360N+V362S/T, B61N, V362N+W364S/T, 3363N+G365S/T, W364N+Q366S/T, G365N+G367S/T, T370N+G372S/T, V376N, f377N+R379S/T, T378N+V380S/T, R379N, V380N-fQ382S/r, Q382N+B84S/T, n83N+E385S/r, W386N+Q388S/T, L387N+K389S/T, L400N-i-R402SyT and combinations hereof. Preferably, the substitution is selected from the group consisting of D289N+G291S/T, ?1290N+A292S/T, G291N, A292N+A294S/T, T293N+L295S/r, S339N+K341S/T, 041N+D343S/T, G342N+S344S/T, D343N+G345S/T, and combinations thereof. More preferably, the substitution is selctedfrom the group consisting of D289N+G291T, 51290N+A292T, G291N, A292N+A294T, T293N+L295T, S339N+K341T, K341N+D343T, 3342N+S344T, D343N-H3345T, and combinations thereof. In addition to a sugar moiety, tlie conjugate according to the aspect of the invention iescribed in the present section may contain additional non-polypeptide moieties, in particular a polymer molecule, as described in the present application, conjugated to one or more ittachment groups present in the polypeptide part of the conjugate. It will bs understood that spy of the amino acid change?, in narticnlar substitutinns. specified in this section can be combined with any of the amino acid changes, in particular substitutions, specified in the other sections herein disclosing specific amino acid changes. For instance, any of the glycosylated variants disclosed in the present section having introduced and/or removed at least one glycosylation site, such as a variant comprising the 5ubstitutions R315N + V317T and/or K143N + N145T, may further be conjugated to a polymer molecule, such as PEG, or any other non-polypeptide moiety. For this purpose the conjugation may be achieved by use of attachment groups already present in the FVTI or FVIIa polypeptide Dr attachment groups may have been introduced and/or removed, in particular such that a total of 1-6, in particular 3-4 or 1, 2,3,4,5, or 6 attachment groups are available for conjugation. Preferably, in a conjugate of the invention wherein the FVH or FVIIa polypeptide comprises two glycosylation sites, the number and molecular weight of the non-polypeptide moiety is chosen so as that the total molecular weight added by the non-polypeptide moiety is in the range of 5-25 kDa, such as in the range of 10-25 KDa, in particular about 5 kDa, about 12 kDa, about 15 kDa or about lOhDsL. An inactive conjugate The conjugates of the invention may be rendered inactive by removing at least one amino acid residue occupying a position selected from tlie group consisting of R152,1153, S344, D242 and H193 of SEQ ID N0:1. The removal may be effected by substitution or deletion of one or more of the above-identified amino acid residues. Preferably, the removal is effected by substitution, in particular by conservative substitution. Accordingly, the inactive F^TI or F\TIa polypeptide used herein may comprise one or more of the following substitutions: R152X, I153X, S344X, D242X or H193X, wherein X is any amino acid residue, preferably one leading to a conservative substitution. For instance, the inactive FVII or FVIIa polypeptide comprises the mutations R152X, wherein X is any amino acid residue other than lysine (since lysine forms part of a protease cleavage site). Other examples of specific substitutions include I153-W/L; S344T/A/G/Y; D242E/A and/or H193R/A. Alternatively, an active FVn or FVIIa polypeptide may be rendered inactive by cai-bamylating the a-amino acid group 1153 or by complexing the polypeptide to a serine proteinase inhibitor. A suitable serine inhibitor protein is, e.g., selected from the group consisting of an organophosphor coumpound, a sulfanylfluoride, a peptide halomethyllcetone, preferably a Dansyl-Phe-Pro-Arg chloromethyllcetone, Dansj'l-Glu-Glu-Arg chlonnethylketone, Dansyl-Phe-Phe-Axg chlormethylketone or a Phe-Phe-Arg chlormethylketone, or an azapeptide. A conjugate may also be rendered inactive by introducing at least one glycosylation site in a position selected so that the subsequent glycosylation inactivates the conjugate. As explained above in the last part of the section entitled "Conjugate of the invention wherein the non-polypeptide motet)' is a sugar moiety" it is preferred that such glycosylation sites are introduced in a position which does not form part of the tissue factor but which forms part of the active site region and the ridge of the active site binding cleft as defined herein. Specific examples of preferred substitutions ar egiven above in the section entitied "Conjugat.e of the invention wherein the non-polypeptide moiety is a sugar moiety". Non-polypeptide moiety of the conjugate of the invention As indicated further above the non-polypeptide moiety of the conjugate of the invention is preferably selected from the group consisting of a polymer molecule, a lipophilic compound, a sugar moiety (by way of in vivo glycosjiation) and an organic derivatizing agent. All of these agents may confer desirable properties to the poh-peptide part of the conjugate, in particular increased functional in vivo half-life and/or increased plasma half-life. The ^ polypeptide part of the conjugate is normally conjugated to only one type of non-polypeptide moiety, but may also be conjugated to two or more different types of non-polypeptide moieties, e.g. to a polymer molecule and a sugar moiety, to a lipophilic group and a sugar moiety, to an organic derivatizing agent and a sugar moiety, to a lipophilic group and a polymer molecule, etc. The conjugation to two or more different non-polypeptide moieties may be done simultaneous or sequentially. Methods of preparing a conjugate of the invention In the following sections "Conjugation to a lipophilic compound", "Conjugation to a polymer molecule", "Conjugation to a sugar moiety" and "Conjugation to an organic derivatizing agent" conjugation to specific types of non-polypeptide moieties is described. In general, a polypeptide conjugate according to the invention may be produced by culturing an appropriate host cell under conditions conducive for the expression of the polypeptide, and recovering the polypeptide, wherein a) the polypeptide comprises at least one N- or O-glycosylation site and the host cell is a eukaryotic host cell capable of in vivo glycosylation, and/or b) the polypeptide is subjected to conjugation to a non-polypeptide moiety in vitro. It will be understood that the conjugation should be designed so as to produce the optimal molecule with respect to the number of non-polypeptide moieties attached, the size and form of such moleculfts (e.g. whether they are linear or branched), and the attachment site(s) in the polypeptide. The molecular weight of the non-polypeptide moiety to be used may e.g. be chosen on the basis of the desired effect to be achieved. For instance, if the primary purpose of the conjugation is to achieve a conjugate having a high molecular weight (e.g. to reduce renal clearance) it is usually desirable to conjugate as few high molecular weight non-polypeptide moieties as possible to obtain the desired molecular weight When a high degree of shielding is desirable this may be obtained by use of a sufficientiy hi^ number of low molecular weight non-polypeptide moieties (e.g. with a molecular weight of from about 300 Da to about 5 IcDa, such as a molecular weight of from 300 Da to 2 kDa) to effectively shield all or most protease cleavage sites or other vulnerable sites of the polypeptide. Conjugation to a lipophilic compound The polypeptide and the lipophilic compound may be conjugated to each other, either directiy or by use of a linker. The lipophilic compound may be a natural compound such as a saturated or unsamrated fatty acid, a fatty acid diketone, a terpene, a prostaglandin, a vitamine, a carotenoide or steroide, or a synthetic compound such as a carbon acid, an alcohol, an amine 3S ana suipnomc acia with one or more alio,'!-, aryl-, alkenyl- or other multiple unsaturated compounds. The conjugation between the polypeptide and the lipophilic compound, optionally through a linker may be done according to methods known in the art, e.g. as described by Bodanszky in Peptide Synthesis, John Wiley, New York, 1976 and in WO 96/12505. Conjugation to a polymer molecule, including conjugation of a polymer molecule to the N-tenninal of the polypeptide The polymer molecule to be coupled to the pohpeptide may be any suitable polymer molecule, such as a natural or synthetic homo-polymer or het&ro-poljTnsr, t>'pically with a molecular weight in the range of about 300-100,000 Da, such as about 500-20,000 Da, more preferably in the range of about 500-15,000 Da, even more preferably in the range of about 2-12 IvDa, such as in the range of about 3-10 IcDa. When the term "about" is used herein in connection with a certain molecular weight, the word "about" indicates an approximate average molecular weight and reflects the fact that there will normally be a certain molecular weight distribution in a given polymer preparation. Examples of homo-polymers include apolyol (i.e. poly-OH), a polyamine. (i.e. poly-NHa) and a polycarboxylic acid (i.e. poly-COOH). A hetero-polymer is a polymer comprising different coupling groups, such as a hydroxyl group and an amine group. Examples of suitable polymer molecules include polymer molecules selected from the group consisting of polyalkydene oxide (PAO), including polyalk}dene glycol (PAG), such as polyethylene glycol (PEG) and polypropylene glycol (PPG), branched PEGs, poly-vinj'l alcohol (PVA), poly-carboxylate, poly-(vinylpyrolidone), polyethylene-co-maleic acid anhydride, polystj'rene-co-maleic acid anhydride, dextran, including carboxymethyl-dextran, or any other biopolymer suitable for reducing immunogenicity and/or increasing functional in vivo half-life and/or serum half-life. Another example of a polymer molecule is human albumin or another abundant plasma protein. Generally, polyallcylene glycol-derived pohoners are biocompatible, non-toxic, non-antigenic, non-immunogenic, have various water solubility properties, and are easily excreted from living organisms. PEG is the preferred pol^aner molecule, since it has only few reactive groups capable of cross-linking compared to, e.g., polysaccharides such as dextran. In particular, mono-functional PEG, e.g. methox}'polyeth}'lene glycol (mPEG), is of interest since its coupling chemistry is relatively simple (only one reactive group is available for conjugating with attachment groups on the pohpeptide). Consequently, the risk of cross-linldng is eliminated. 3^ the resulting polj'peptide conjugates are more uuuaus molecules with the polypeptide is easier to control. To effect covalent attachment of the polymer molecule(s) to the polypeptide, the hydroxyl end groups of the polymer molecule must be provided in activated form, i.e. with reactive functional groups (examples of which include primary amino groups, hydrazide (HZ), thiol, succinate (SUC), succinimidyl succinate (SS), succinimidyl succinamide (SSA), succinimidyl proprionate (SPA), succinimidy carbox}'methylate (SCM), benzotriazole carbonate (BTC), N-hydroxysuccinimide (NHS), aldehyde, nitrophenylcarbonate (NPC), and tresylate (TRES)). Suitable activated polymer molecules are commercially available, e.g. from Shearwater Polymers, Inc., Huntsville, AL, USA, or from PolyMASC Pharmaceuticals pic, UTL Alternatively, the polymer molecules can be activated by conventional methods known in the art, e.g. as disclosed in WO 90/13540. Specific examples of activated linear or branched polymer molecules for use in the present invention are described in the Shearwater Polymers, Inc. 1997 and 2000 Catalogs (Functionalized Biocompatible Polymers for Research and pharmaceuticals, Polyethylene Glycol and Derivatives, incorporated herein by reference). Specific examples of activated PEG polymers include the following linear PEGs: NHS-PEG (e.g. SPA-PEG, SSPA-PEG, SBA-PEG, SS-PEG, SSA-PEG, SC-PEG, SG-PEG, and SCM-PEG), and NCR-PEG, BTC-PEG, EPOX-PEG, NCO-PEG, NPC-PEG, CDI-PEG, ALD-PEG, TRES-PEG, VS-PEG. lODO-PEG, and MAL-PEG, and branched PEGs such as PEG2-NHS and those disclosed in US 5,932,462 and US 5,643,575, both of which are incorporated herein by reference. Furthermore, the following publications, incorporated herein by reference, disclose useful polymer molecules and/or PEGylation chemistries: US 5,824,778, US 5,476,653, WO 97/32607, EP 229,108, EP 402,378, US 4,902,502, US 5,281,698, US 5,122,614, US 5,219,564. WO 92/16555, WO 94/04193, WO 94/14758, WO 94/17039, WO 94/18247, WO 94/28024, WO 95/00162, WO 95/11924, WO95/13090, WO 95/33490, WO 96/00080, WO 97/18832, WO 98/41562, WO 98/48837, WO 99/32134, WO 99/32139, WO 99/32140, WO 96/40791, WO 98/32466, WO 95/06058, EP 439 508, WO 97/03106, WO 96/21469, WO 95/13312, EP 921 131, US 5,736,625, WO 98/05363, EP 809 996, US 5,629,384, WO 96/41813, WO 96/07670, US 5,473,034, US 5,516,673, EP 605 963, US 5,382,657, EP 510 356, EP 400 472, EP 183 503 and EP 154 316. The conjugation of the polypeptide and the activated polymer molecules is conducted by use of any conventional method, e.g. as described in the following references (which also describe suitable methods for activation of polymer molecules); R.F. Taylor, (1991), "Protein immobilisation. Fundamental and applications". Marcel Dekker, N.Y.; S.S. Wong, (1992), CncmisxT}' 01 rrotem uonjuganon ana crossiinKing', UKCrress,rjonaa, U^A; It will be understood that the PEGylation is designed so as to produce the optimal molecule with respect to the number of PEG molecules attached, the size and form of such molecules (e.g. whether they are lineai' or branched), and the attachment site(s) in the,,,, polypeptide. The molecular weight of the polymer to be used may e.g. be chosen on the basis of the desired effect to be achieved. For instance, if the primary purpose of the conjugation is to achieve a conjugate having a high molecular weight (e.g. to reduce renal clearance) it is usually desirable to conjugate as few high molecular weight poisoner molecules as possible to obtain the desired molecular weight. When a high degree of shielding is desirable this may be obtained by use of a sufficiently high number of low molecular weight pol}'mer molecules (e.g. with a molecular weight of from about 300 Da to about 5 IcDa) to effectively shield all or most protease cleavage sites or other vulnerable sites of the pol^'peptide. For instance, 2-8, such as 3-6 such poljoners may be used. In connection with conjugation to only a single attachment group on the protein (e.g. the N-tetminal amino group), it may be advantageous tliat the polymer molecule, which may be linear or branched, has a high molecular weight, preferably about 10-25 IcDa, such as about 15-25 IcDa, e.g. about 20 kDa. Normally, the poljoner conjugation is performed under conditions aimed at reacting as many of the available polymer attachment groups with pol^Tner molecules. This is achieved by means of a suitable molar excess of the polymer relative to the pol>'peptide. Typically, the molar ratios of activated polymer molecules to polj'peptide are up to about 1000-1, such as up to about 200-1, or up to about 100-1. In some cases the ration may be somewhat lower, however, such as up to about 50-1,10-1 or 5-1 in order to obtain optimal reaction. ^6 It is also contemplated according to the invention to couple the polymer molecules to he polypeptide through a linker. Suitable linkers are well known to the skilled person. A referred example is cyanuric chloride (Abuchowski et al., (1977), J. Biol. Chem., 252, 5578-3581; US 4,179,337; Shafer et al, (1986), J. Polym. Sci. Polym. Chem. Ed., 24,375-378). Subsequent to the conjugation, residual activated polymer molecules are blocked iccording to methods known in the art, e.g. by addition of primary amine to the reaction nixture, and the resulting inactivated polymer molecules are removed by a suitable method. It will be understood that depending on the circumstances, e.g. the amino acid sequence of the polj'peptide, the nature of the activated PEG compound being used and the specific PEGylation conditions, including the molar ratio of PEG to polypeptide, varying degrees of PEGylation may be obtained, with a higher degree of PEGylation generally being Dbtained with a higher ratio of PEG to polypeptide. The PEGylated polypeptides resulting from any given PEGylation process will, however, normally comprise a stochastic distribution of polypeptide conjugates having slightly different degrees of PEGylation. In an interesting embodiment of the invention the polypeptide conjugate of the invention comprises a polymer molecule covalently attached to the Al N-terminal of the wild 'ype FVU or FVIIa polypeptide shown in SEQ ID N0:1, where said polymer molecule is the Dnly polymer molecule attached to the polypeptide. Preferably, such polypeptide conjugates are Dnss, which comprise a single PEG mol'°'-^'i1^- attachftd to the N-tfirmina1 of the oolvoeotide and no other PEG molecules. In particular, a linear or branched PEG molecule with a molecular weight of at least about 5 kDa, in particular about 10-25 ld)a, such as about 15-25 kDa, e.g. about 20 IcDa is preferred. The polypeptide conjugate according to this embodiment may further comprise one or more sugar moieties attached to an N-linked or 0-linked glycosylation site of the polypeptide or sugar moieties attached by in vitro glycosylation. In a further interesting embodiment of the invention the polypeptide conjugate of the invention comprises poljmer molecules covalently attached to the Al N-terminal and to the 1153 N-terminal of the wild type FVIIa polypeptide shown in SEQ ID N0:1, where said polymer molecules are the only polymer molecules attached to the polypeptide. Preferably, such polypeptide conjugates are ones, which comprise a PEG molecule attached to both of the N-terminals of FVHa and no other PEG molecules, hi particular, linear or branched PEG molecules with a molecular weight of at least about 5 kDa, in particular about 10-25 kDa, such as about 15-25 kDa, e.g. about 20 kDa are preferred. The polypeptide conjugate according to this embodiment may further comprise one or more sugar moieties attached to an N-linked or 37 0-linked glycosylation site of the polypeptide or sugar moieties attached by in vitro glycosylation. One preferred method for selectively coupling polymer molecules, such as PEG molecules, to the N-terminal of the polypeptide is the method disclosed in US 5,985,265. This method involves reductive alkylation (reaction of the N-terminal amino group of the polypeptide with an aldehyde-containing polypeptide, such as aldehylde-PEG, in the presence of a reducing agent, such as NaCNBHs). This method exploits differential reactivity of different types of primary amino groups (lysine versus N-terminal) available for derivatization in the polj'peptide, thereby achieving substantially selective derivatization of the polypeptide at the N-terminus with a carbonyl group-containing polymer molecule, such as aldehyde-PEG. The reaction is performed at a pH which allows one to take advantage of the pKa differences between the e-amino groups of the lysine residues and that of the a-amino group of the N-terminal residue of the polypeptide. In order to achieve this differential reactivity, the reaction is typically carried at slightly acidic conditions. Specific examples of suitable pH ranges include pH 4.5-7, such as pH 4.5-6, e.g. pH 5-6, in particular about pH 5, In another specific embodiment, the polypeptide conjugate of the invention comprises a PEG molecule attached to each of the lysine residues in the polypeptide available for PEGylation, in particular a linear or branched PEG molecule, e.g. with a molecular weight of about 1-15 IdDa, typically about 2-12 IcDa, such as about 3-10 IdDa, e.g. about 5 or 6 kDa. In yet another embodiment, the polypeptide conjugate of the invention comprises a PEG molecule attached to each of the lysine residues in the polypeptide available for PEGylation, and in addition to the N-terminal amino acid residue of the polypeptide. Covalent in vitro coupling of carbohydrate moieties (such as dextran) to amino acid residues of the polypeptide may also be used, e.g. as described, for example in WO 87/05330 and in Aplin etl al., ORG Crit Rev. Biochem, pp. 259-306,1981. The in vitro coupling of carbohydrate moieties or PEG to protein- and peptide-bound Gin-residues can be carried out by transglutaminases (TGases). Transglutaminases catalyse the transfer of donor amine-groups to protein- and peptide-bound Gin-residues in a so-called cross-linldng reaction. The donor-amine groups can be protein- or peptide-bound , such as the £-amino-group in Lys-residues or it can be part of a small or large organic molecule. An example of a small organic molecule functioning as amino-donor in TGase-catalysed cross-linking is putrescine (1,4-diaminobutane). An example of a larger organic molecule functioning as amino-donor in TGase-catalysed cross-linking is an amine-containing PEG (Sato et al., 1996, Biochemistry 35,13072-13080). 3? TGases, in general, are highly specific enzymes, and not every Ciin-residues exposed on the surface of a protein is accessible to TGase-catalysed cross-linldng to amino-containing substances. On the contrary, only few Gin-residues are naturally functioning as TGase substrates but the exact parameters governing which Gin-residues are good TGase substrates remain unknown. Thus, in order to render a protein susceptible to TGase-catalysed cross--Unking reactions it is often a prerequisite at convenient positions to add stretches of amino acid sequence known to function very well as TGase substrates. Several amino acid sequences are known to be or to contain excellent natural TGase substrates e.g. substance P, elafin, fibrinogen, fibronectin, a2-plasmin inhibitor, a-caseins, and ^-caseins. Coupling to a sugar moiety In order to achieve in vivo glycosylation of a FVH molecule comprising one or more glycosylation sites the nucleotide sequence encoding the polypeptide must be inserted in a glycosylating, eucaryotic expression host. The expression host cell may be selected from fungal (filamentous fungal or yeast), insect or animal cells or from transgenic plant cells. In one embodiment the host cell is a mammalian cell, such as a CHO cell, BHK or HEK, e.g. HEK 293, cell, or an insect cell, such as an SF9 cell, or a yeast cell, e.g. 5". cerevisiae or Pichia pastoris, or any of the host cells mentioned hereinafter. Coupling to an organic derivatizing agent Covalent modification of the polypeptide may be performed by reacting one or more attachment groups of the polypeptide with an organic derivatizing agent. Suitable derivatizing agents and methods are well known in the art. For example, cysteinyl residues most commonly are reacted with a-haloacetates (and corresponding amines), such as chloroacetic acid or chloroacetamide, to give carbox}'methyl or carboxyamidomethyl derivatives. Cysteinyl residues also are derivatized by reaction with bromotrifluoroacetone, a-bromo-P-(4-imidozoyl)propionic acid, chloroacetyl phosphate, N-aUcylmaleimides, 3-nitro-2-pyridyl disulfide, methyl 2-pyridyl disulfide, p-chloromercuribenzoate, 2-chloromercuri-4-nitrophenol, or chloro-7-nitrobenzo-2-oxa-l,3-dia2ole. Histidyl residues are derivatized by reaction with diethylpyrocarbonateat pH 5.5-7.0 because this agent is relatively specific for the histidyl side chain. Para-bromophenacyl bromide also is useful. The reaction is preferably performed in 0.1 M sodium cacodylate at pH 6.0. Lysinyl and amino terminal residues are reacted v^dth succinic or other carbox^'lic acid anhydrides. Derivatization with these agents has the effect of reversing the charge of the lysinyl 3^ residues. Other suitable reagents for derivatizing a-amino-containing residues include imidoesters such as methyl picohnimidate, pyridoxal phosphate, pyridoxal, chloroborohydride, Irinitrobenzenesulfonic acid, 0-methyIisourea, 2,4-pentanedione and transaminase-catalyzed reaction with glyox5'late. Ai'ginyl residues are modified by reaction with one or several conventional reagents, among them phenylglyoxal, 2,3-butanedione, 1,2-cyclohexanedione, and ninhydrin. Derivatization of arginine residues requires that the reaction be performed in alkaline conditions because of tlie high pKa of the guanidine functional group. Furthermore, these reagents may react with the groups of lysine as well as the arginine guanidino group. Carboxyl side groups (aspartyl or glutamyl) are selectively modified by reaction with carbodiimides (R-N=C=N-R'), where R and R' are different allc}4 groups, such as l-cyclohexyl-3-(2-morphohnyl-4-ethyl) carbodiimide or l-ethyl-3-(4-azonia-4,4-dimethylpentj'l) carbodiimide. Furthermore, aspartyl and glutamyl residues are converted to asparaginyl and glutaminyl residues by reaction with ammonium ions. Blocking of functional site It has been reported that excessive polymer conjugation can lead to a loss of activity of the polypeptide to which the non-polypeptide moiety is conjugated. This problem can be eliminated, e.g., by removal of attachment groups located at the functional site or by reversible blocldng the functional site prior to conjugation so that the functional site is blocked during conjugation. The latter strategy constitutes further embodiments of the invention (the first strategy being exemplified further above, e.g. by removal of lysine residues which may be located close to the functional site). More specifically, according to the second strategy the conjugation betvi'een the polypeptide and the non-pol)peptide moiety is conducted under conditions where the functional site of the polypeptide is blocked by a helper molecule e.g. tissue factor capable of binding to the functional site of the polj'peptide or a serine protease inhibitor. Preferably, the helper molecule is one, which specifically recognizes a functional site of the polypeptide, such as a receptor, in particular tissue factor, either full length or a suitably truncated form of tissue factor or two molecules, one being tissue factor the other one being a peptide or peptide inhibitor binding to and thus protecting the area around the catalytic triad (preferably defined as amino acid residues viithin 10 A of any atom in the catalytic triad). ifO Alternatively, the helper molecule may be an antibody, in particular a monoclonal antibody recognizing the FVn polypeptide. In particular, liie helper molecule may be a neutralizing monoclonal antibody. The polypeptide is allowed to interact with the helper molecule before effecting conjugation. This ensures that the functional site of the polypeptide is shielded or protected and consequently unavailable for derivatization by the non-polypeptide moiety such, as a polymer. Following its elution from the helper molecule, the conjugate between the non-polypeptide moiety and the polypeptide can be recovered with at least a partially preserved functional site. The subsequent conjugation of the polypeptide having a blocked functional site to a polymer, a lipophilic compound, a sugar moiety, an organic deiivatizing agent or any other compound is conducted in the normal way, e.g. as described in the sections above entitled "Conjugation to ....". Irrespectively of tlie nature of the helper molecule to be used to shield the functional site of the polypeptide from conjugation, it is desirable that the helper molecule is free from or comprises only few attachment groups for the non-polypeptide moiety of choice in part(s) of the molecule, where the conjugation to such groups will hamper the desorption of the conjugated polypeptide from the helper molecule. Hereby, selective conjugation to attachment groups present in non-shielded parts of the pol3'peptide can be obtained and it is possible to reuse the helper molecule for repeated cycles of conjugation. For instance, if the non-polypeptide moiety is a polymer molecule such as PEG, which has the epsilon amino group of a lysine or N-teiminal amino acid residue as an attachment group, it is desirable that the helper molecule is substantially free from conjugatable epsilon amino groups, preferably free from any epsilon amino groups. Accordingly, in a preferred embodiment the helper molecule is a protein or peptide capable of binding to the functional site of the polypeptide, which protein or peptide is free from any conjugatable attachment groups for the non-polypeptide moiety of choice. In a further embodiment the helper molecule is first covalently linked to a solid phase such as column packing materials, for instance Sephadex or agarose beads, or a surface, e.g. reaction vessel. Subsequently, the polypeptide is loaded onto the column material carrying the helper molecule and conjugation carried out according to methods known in the art, e.g. as described in the sections above entitied "Conjugation to ....". This procedure allows the polypeptide conjugate to be separated from the helper molecule by elution. The polypeptide conjugate is eluted by conventional techniques under physico-chemical conditions that do not lead to a substantive degradation of the polypeptide conjugate. The fluid phase containing the pol}'peptide conjugate is separated from the solid phase to which the helper molecule remains ^l covalently linlced. The separation can be achieved in other ways: For instance, the helper molecule may be derivatised with a second molecule (e.g. biotin) that can be recognized by a specific binder (e.g. streptavidin). The specific binder may be linked to a solid phase thereby allowing the separation of the polypeptide conjugate from the helper molecule-second molecule complex through passage over a second helper-solid phase column which will retain, upon subsequent elution, the helper molecule-second molecule complex, but not the polypeptide conjugate. The po]3'peptide conjugate may be released from the helper molecule in any appropriate fashion. Deprotection may be achieved by providing conditions in which the helper molecule dissociates from the functional site of the FVH to which it is bound. For instance, a complex between an antibody to which a polymer is conjugated and an anti-idiotypic antibody can be dissociated by adjusting the pH to an acid or alkaline pH. Even more preferred is the use of a conformation specific antibody that recognizes a Ca^"*" specific conformation of FVH and consequently can be eluted with EDTA under mild conditions. Attachment of sej-ine protease inhibitor Attachment of a serine protease inhibitor can be performed in accordance vi'ith the method described in WO 96/12800. Conjugation of a tagged polypeptide In an alternative embodiment the polypeptide is expressed as a fusion protein with a tag, i.e. an amino acid sequence or peptide stretch made up of tj'pically 1-30, such as 1-20 amino acid residues. Besides allowing for fast and easy purification, the tag is a convenient tool for achieving conjugation between the tagged polypeptide and the non-polypeptide moiety. In particular, the tag may be used for achieving conjugation in microtiter plates or other carriers, such as paramagnetic beads, to which the tagged polypeptide can be immobilised via the tag. The conjugation to the tagged polypeptide in, e.g., microtiter plates has the advantage that the tagged polj'peptide can be immobilised in the microtiter plates directly from the culture broth (in principle without any purification) and subjected to conjugation. Thereby, the total number of process steps (from expression to conjugation) can be reduced. Furthermore, the tag may function as a spacer molecule, ensuring an improved accessibility to the immobilised polypeptide to be conjugated. The conjugation using a tagged polxpeptide may be to any of the non-polypeptide moieties disclosed herein, e.g. to a poljmer molecule such as PEG. The identit}' of the specific tag to be used is not critical as long as the tag is capable of being expressed v»'ith the poh'peptide and is capable of being immobilised on a suitable surface or carrier material. A number of suitable tags are commercially available, e.g. from Unizyme Laboratories, Denmark. For instance, the tag may consist of any of the following sequences: His-His-His-His-His-His Met-Lys-IEs-His-His-His-His-]ffis Met-Lys-His-His-Ala-His-His-Gln-His-His Met-Lys-His-Gln-His-Gb-His-Gln-His-Gln-His-Gln-His-Gln Met-Lys-His-Gln-His-Gb-His-Gln-His-Gln-His-Gln-His-Gln-Gln or any of the following: EQKU SEEDL (a C-terminal tag described in Mol. Cell. Biol. 5:3610-16, 1985) DYKDDDDK (a C- or N-terminal tag) YPYDVPDYA Antibodies against the above tags are commercially available, e.g. from ADI, Aves Lab and Research Diagnostics. The subsequent cleavage of the tag from the polypeptide may be achieved by use of ■ commercially available enzymes. Methods of preparing a polypeptide of the invention or the polypeptide of the conjugate qftJie invention The polvpentide of A nucleotide sequence encoding a polypeptide or the polypeptide part of a conjugate of the invention may be constructed by isolating or synthesizing a nucleotide sequence encoding the parent FVH, such as hFVn with the amino acid sequence shown in SEQ ID N0:1 and then changing the nucleotide sequence so as to effect introduction (i.e. insertion or substitution) or removal (i.e. deletion or substitution) of the relevant amino acid residue(s). TTie nucleotide sequence is conveniently modified by site-directed mutagenesis in accordance with conventional methods. Alternatively, the nucleotide sequence is prepared by chemical synthesis, e.g. by using an oligonucleotide synthesizer, wherein oligonucleotides are designed based on the amino acid sequence of the desired polypeptide, and preferably selecting ^3 those codons that are faA'ored in the host cell in which the recombinant polypeptide will be produced. For example, several small oligonucleotides coding for portions of the desired polypeptide may be synthesized and assembled by PCR, ligation or ligation chain reaction (LCR) (Barany, PNAS 88:189-193,1991). The individual oligonucleotides typically contain 5' or 3' overhangs for complementary assembly. Alternative nucleotide sequence modification methods are available for producing pol5'peptide variants for high throughput screening, for instance methods which involve homologous cross-over such as disclosed in US 5,093,257, and methods which involve gene shuffling, i.e. recombination between two or more homologous nucleotide sequences resulting in new nucleotide sequences having a number of nucleotide alterations when compared to the starting nucleotide sequences. Gene shuffling (also Icnown as DNA shufOing) involves one or more cycles of random fragmentation and reassembly of the nucleotide sequences, followed by screening to select nucleotide sequences encoding polypeptides with desired properties. In order for homology-based nucleic acid shuffling to talce place, the relevant parts of the nucleotide sequences are preferably at least 50% identical, such as at least 60% identical, more preferably at least 70% identical, such as at least 80% identical. The recombination can be performed in vitro or in vivo. Examples of suitable in vitro gene shuffling methods are disclosed by Stemmer et al. (1994), Proc. Natl. Acad. Sci. USA; vol. 91, pp. 10747-10751; Stemmer (1994), Nature, vol. 370, pp. 389-391; Smith (1994), Nature vol. 370, pp. 324-325; Zhao et al., Nat. Biotechnol. 1998, Mar; 16(3): 258-61; Zhao H. and Arnold, FB, Nucleic Acids Research, 1997, Vol. 25. No. 6 pp. 1307-1308; Shao et al.. Nucleic Acids Research 1998, Jan 15; 26(2): pp. 681-83; and WO 95/17413. An example of a suitable in vivo shuffling method is disclosed in WO 97/07205. Other techniques for mutagenesis of nucleic acid sequences by in vitro or in vivo recombination are disclosed e.g. in WO 97/20078 and US 5,837,458. Examples of specific shuffling techniques include "family shuffling", "synthetic shuffling" and "wz silico shuffling". Family shuffling involves subjecting a family of homologous genes from different species to one or more cycles of shuffling and subsequent screening or selection. Family shuffling techniques are disclosed e.g. by Crameri et al. (1998), Nature, vol. 391, pp. 288-291; Christians et al. (1999), Nature Biotechnology, vol, 17, pp. 259-264; Chang et al. (1999), Nature Biotechnology, vol. 17, pp. 793-797; and Ness et al. (1999), Nature Biotechnology, vol. 17,893-896. ^if Synthetic shuffling involves providing libraries of overlapping s}'nthetic oligonucleotides based e.g. on a sequence alignment of homologous genes of interest. The synthetically generated oligonucleotides are recombined, and the resulting recombinant nucleic acid sequences are screened and if desired used for further shuffling cycles. Synthetic shuffling techniques are disclosed in WO 00/42561. In silico shuffling refers to a DNA shuffling procedure, which is performed or modelled using a computer system, thereby partly or entirely avoiding the need for physically manipulating nucleic acids. Techniques for in silico shuffling are disclosed in WO 00/42560. Once assembled (by synthesis, site-directed mutagenesis or another method), the nucleotide sequence encoding tlie polypeptide is inserted into a recombinant vector and operably Knked to control sequences necessary for expression of the FVII in the desired transformed host cell. It should of course be understood that not all vectors and expression control sequences function equally well to express the nucleotide sequence encoding a polypeptide described herein. Neither will all hosts function equally well with the ssmt expression system. However, one of skill in the art may make a selection among these vectors, expression control sequences and hosts without undue experimentation. For example, in selecting a vector, the host must be considered because the vector must replicate in it or be able to integrate into the chromosome. The vector's cnnv number, the abilitv to control that conv number, and the expression of anv Other proteins encoded by the vector, such as antibiotic markers, should also be considered. In selecting an expression control sequence, a variety of factors should also be considered. These include, for example, the relative strength of the sequence, its controllability, and its compatibility with the nucleotide sequence encoding the polypeptide, particularly as regards potential secondary structures. Hosts should be selected by consideration of their compatibiUty with the chosen vector, the toxicity of the product coded for by the nucleotide sequence, their secretion characteristics, their ability to fold the polypeptide correctly, their fermentation or culture requirements, and the ease of purification of the products coded for by the nucleotide sequence. The recombinant vector may be an autonomously replicating vector, i.e. a vector, which exists as an extrachromosomal entity, the replication of which is independent of chromosomal replication, e.g. a plasmid. Alternatively, the vector is one which, when introduced into a host cell, is integrated into the host cell genome and replicated together with the chromosome(s) into which it has been integrated. H^ The \'ector is preferably an expression vector, in which the nucleotide sequence encoding the polypeptide of the invention is operably linked to additional segments required for transcription of the nucleotide sequence. The vector is typically derived from plasmid or viral DNA. A number of suitable expression vectors for expression in the host cells mentioned herein ai-e commercially available or described in the literature. Useful expression vectors for eukaryotic hosts, include, for example, vectors comprising expression control sequences from SV40, bovine papilloma virus, adeno^'irus and cytomegalovirus. Specific vectors are, e.g., pCDNA3.1(+)\H3'g (Invitrogen, Carlsbad, CA, USA) and pCI-neo (Stratagene, La Jola, CA, USA), Useful expression vectors for yeast cells include the 2}i plasmid-and derivatives thereof, the POTl vector (US 4,931,373), the pJS037 vector described in Okicels, Ann. New York Acad. Sci. 782, 202-207,1996, and pPICZ A, B or C (Invitrogen). Useful vectors for insect cells include pVL941, pBG311 (Gate et al, "Isolation of the Bovine and Human Genes for MuUerian Inhibiting Substance And Expression of the Human Gene In Animal Cells", Cell, 45, pp. 685-98 (1986), pBluebac 4.5 and pMelbac (both available from Invitrogen). Useful' expression vectors for bacterial hosts include Icnown bacterial plasmids, such as plasmids from E. coli, including pBR322, pET3a and pET12a (both from Novagen Inc., WI, USA), wider host range plasmids, such as RP4, phage DNAs, e.g., the numerous derivatives of phage lambda, e.g., } Other vectors for use in this invention include those that allow the nucleotide sequence encoding the polypeptide to be amplified in copy number. Such amplifiable vectors are well Icnown in the art. They include, for example, vectors able to be amplified by DHFR amplification (see, e.g., Kaufman, U.S. Pat. No. 4,470,461, Kaufman and Sharp, "Construction Of A Modular Dihydrafolate Reductase cDNA Gene: Analysis Of Signals Utilized For Efficient Expression", Mol. Cell. Biol., 2, pp. 1304-19 (1982)) and glutamine s^Tithetase ("GS") amplification (see, e.g., US 5,122,464 and HP 338,841). The recombinant vector may further comprise a DNA sequence enabling the vector to replicate in the host cell in question. An example of such a sequence (when the host cell is a mammalian cell) is the SV40 origin of replication. 'VvTien the host cell is a yeast cell, suitable sequences enabling the vector to replicate are the yeast plasmid 2ji replication genes REP 1-3 and origin of replication. The vector may also comprise a selectable marker, e.g. a gene the product of which complements a defect in the host cell, such as the gene coding for dihydrofolate reductase ^h (DHFR) or the Schizosaccharomyces pombe TPI gene (described by P.R. Russell, Gene 40, 1985, pp. 125-130), or one which confers resistance to a drug, e.g. ampicillin, kanamycin, tetracyclin, chloramphenicol, neomycin, hygromycin or methotrexate. For Saccharonryces cerevisiae, selectable markers include ura3 and leu2. For filamentous fungi, selectable markers include amdS, pyrG, arcB, nioD and sC. The term "control sequences" is defined herein to include all components, which are necessary or advantageous for the expression of the polypeptide of the invention. Each control sequence may be native or foreign to the nucleic acid sequence encoding the polypeptide. Such control sequences include, but are not limited to, a leader sequence, polyadenylation sequence, propeptide sequence, promoter, enhancer or upstream activating sequence, signal peptide sequence, and transcription terminator. At a minimum, the control sequences include a promoter. A wide variety of expression control sequences may be used in the present invention. Such useful expression control sequences include the expression control sequences associated with structural genes of the foregoing expression vectors as well as any sequence known to control the expression of genes of prokaryotic or eukaryotic cells or their viruses, and various combinations thereof. Examples of suitable control sequences for directing transcription in mammalian cells include the early and late promoters of S V40 and adenovirus, e.g. the adenovirus 2 major late promoter, the MT-1 (metallothionein gene) promoter, the human cytomegalovirus immediate-early gene promoter (CMV), the human elongation factor la (EF-loc) promoter, the Drosophila minimal heat shock protein 70 promoter, the Rous Sarcoma Virus (RSV) promoter, the human ubiquitin C (UbC) promoter, the human grovv^h hormone terminator, SV40 or adenovirus Elb region polyadenylation signals and the Kozalc consensus sequence (Kozak, M. JMol Biol 1987 Aug 20;196(4):947-50). In order to improve expression in mammalian cells a synthetic intron may be inserted in the 5' untranslated region of the nucleotide sequence encoding the polypeptide. An example of a s>'nthetic intron is the synthetic intron from the plasmid pQ-Neo (available from Promega Corporation, WI, USA). Examples of suitable control sequences for directing transcription in insect cells include the polyhedrin promoter, the PIO promoter, the AutograpJxa califomica polyhedrosis virus basic protein promoter, the baculovirus immediate early gene 1 promoter and the baculovirus 39K delayed-early gene promoter, and the S V40 polyadenylation sequence. c^l - Examples of suitable control sequences for use in yeast host cells include the promoters of the yeast a-mating system, the yeast triose phosphate isomerase (TPI) promoter, promoters from yeast glycolytic genes or alcohol dehydrogenase genes, the AI)H2-4c promoter, and the inducible GAL promoter. Examples of suitable control sequences for use in filamentous fungal host cells include the ADH3 promoter and terminator, a promoter derived from the genes encoding Aspergillus oryzae TAKA. amylase triose phosphate isomerase or alicaline protease, an A. niger a-amylase, A. niger or A. nidulans glucoamylase, A. nidulans acetamidase, Rhizomucor miehei aspartic proteinase or lipase, the TPIl terminator and the ADH3 terminator. Examples of suitable control sequences for use in bacterial host cells include promoters of the lac system, the trp system, the TAC or TRC system, and the major promoter regions of phage lambda. The presence or absence of a signal peptide will, e.g., depend on the expression host cell used for the production of the polypeptide to be expressed (whether it is an intracellular or extracellular polypeptide) and whether it is desirable to obtain secretion. For use in filamentous fungi, the signal peptide may conveniently be derived from a gene encoding an Aspergillus sp. amylase or glucoamylase, a gene encoding a Rhizomucor miehei lipase or protease or a Humicola lanuginosa lipase. The signal peptide is preferably derived from a gene encoding A. oryzae TAKA amylase, A. niger neutral a-amylase, A, niger acid-stable amylase, or A. niger glucoamylase. For use in insect cells, the signal peptide may conveniently be derived from an insect gene (cf. WO 90/05783), such as the Lepidopteran maiiduca sexta adipolcinetic hormone precursor, (cf. US 5,023,328), the honeybee meiittin (Invitrogen), ecdysteroid UDPglucosyltransferase (egt) (Murphy et al., Protein Expression and Purification 4, 349-357 (1993) or human pancreatic lipase (hpl) (Methods in Enzymology 284, pp. 262-272,1997). A preferred signal peptide for use in mammalian cells is that of hFVn or the murine Ig kappa light chain signal peptide (Coloma, M (1992) J. Imm. Methods 152:89-104). For use in yeast cells suitable signal peptides have been found to be the a-factor signal peptide from S. cereviciae (cf, US 4,870,008), a modified carboxypeptidase signal peptide (cf. L.A. Vails et al, Cell 48,1987, pp. 887-897), the yeast BARl signal peptide (cf. WO 87/02670), the yeast aspartic protease 3 (YAPS) signal peptide (cf. M. Egel-Mitani et al. Yeast 6, 1990, pp. 127-137), and the synthetic r leader sequence TA57 (W098/32867). For use in E. coli cells a suitable signal peptide have been found to be the signal peptide ompA (EP5S1821). The nucleotide sequence of the invention encoding aFVTl polypeptide, whether prepared by site-directed mutagenesis, synthesis, PCR or other methods, may optionally include J?? . a nucleotide sequence that encode a signal peptide. The signal peptide is present when the polypeptide is to be secreted from the cells in which it is expressed. Such signal peptide, if present, should be one recognized by the cell chosen for expression of the polypeptide. The signal peptide may be homologous (e.g. be that normally associated with hFVII) or heterologous (i.e. originating from another source than hFVII) to the polypeptide or may be homologous or heterologous to the host cell, i.e. be a signal peptide normally expressed from the host cell or one which is not normally expressed from the host ceU. Accordingly, the signal peptide may be prokaryotic, e.g. derived from a bacterium such as E. coli, or eukaryotic, e.g. derived from a mammalian, or insect or yeast cell. Any suitable host may be used to produce the polypeptide or polypeptide part of the conjugate of the invention, including bacteria, fungi (including yeasts), plant, insect, mammal, or other appropriate animal cells or ceU lines, as well as transgenic animals or plants. Examples of bacterial host cells include grampositive bacteria such as strains of Bacillus, e.g. B. brevis or 5. subtilis, Pseudomonas or Streptomyces, or gramnegative bacteria, such as strains of £. coli. The introduction of a vector into a bacterial host cell may, for instance, be effected by protoplast transformation (see, e.g., Chang and Cohen, 1979, Molecular General Genetics 168: 111-115), using competent cells (see, e.g., Young and Spizizin, 1961, Journal of Bacteriology 81: 823-829, or Dubnau and Davidoff-Abelson, 1971, Journal of Molecular Biology 56: 209-221), electroporation (see, e.g., Shigekawa and Dower, 1988, Biotechniques 6: 742-751), or conjugation (see, e.g., Koehler and Thome, 1987, Journal of Bacteriology 169: 5771-5278). Examples of suitable filamentous fungal host cells include strains of Aspergillus, e.g. A. oryzae, A. niger, or A nidulans, Fusarium or Trichodenna. Fungal cells may be transformed by a process involving protoplast formation, transformation of the protoplasts, and regeneration of the cell wall in a manner known per se. Suitable procedures for transformation of Aspergillus host cells are described in EP 238 023 and US 5,679,543. Suitable methods for transforming Fusarium species are described by Malardier et al., 1989, Gene 78: 147-156 and WO 96/00787. Examples of suitable yeast host cells include strains of Saccharomyces, e.g. S. cerevisiae, Schizosaccliaromyces, Klyverojnyces, Pichia, such as P. pastoris or P. metlianolica, Hansenula, such as H. Polymorplia or Yarrowia. Yeast may be transformed using the procedures described by Becker and Guarente, In Abelson, J.N. and Simon, M.L, editors, Guide to Yeast Genetics and Molecular Biology, Methods in Enzymology, Volume 194, pp 182-187, Academic Press, Inc., New York; Ito et al, 1983, Journal of Bacteriology 153: 163; Hinnen et al, 1978, Proceedings of the National Academy of Sciences USA 75: 1920: and as disclosed by Clontech Laboratories, Inc. Palo Alto, CA, USA (in the product protocol for the Yeastmaker"^^ Yeast Transformation System Kit). Examples of suitable insect host cells include a Lepidoptora cell line, such as Spodoptera fiiigiperda (Si9 or Sf21) or Trichoplusioa ni cells (High Five) (US 5,077,214). Transformation of insect cells and production of heterologous polypeptides therein may be performed as described by In^'itrogen. Examples of suitable mammalian host cells include Chinese hamster ovary (CHO) cell lines, (e.g. CHO-Kl; ATCC CCL-61), Green Monlcey cell lines (COS) (e.g. COS 1 (ATCC CRL-1650), COS 7 (ATCC CRL-1651)); mouse cells (e.g. NS/0), Baby Hamster Kidney (BHK) ct]l lines (e.g. ATCC CRL-1632 or ATCC CCL-10), and human cells (e.g. HEK 293 (ATCC CRL-1573)), as well as plant cells in tissue culture. Additional suitable cell lines are known in the art and available from pubhc depositories such as the American Type Culture Collection, Rock\'ille, Marj'land. Also, the mammalian cell, such as a CHO cell, may be modified to express sial3'1 transferase, e.g. 1,6-sialyltransferase, e.g. as described in US 5,047,335, in order to pro^dde improved glycosylation of the FVn or FVIIa polypeptide. in order to increase secretion it may be of particular interest to produce the polypeptide of the invention together with an endoprotease, in particular a PACE (Paired basic amino acid converting enz}'me) (e.g. as described in US 5,986,079), such as a Kex2 endoprotease (e.g. as described in WO 00/28065). Methods for introducing exogeneous DNA into mammalian host cells include calcium phosphate-mediated transfection, electroporation, DEAE-dextran mediated transfection, hposome-mediated transfection, viral vectors and the transfection method described by life Tecimologies Ltd, Paisley, UK using Lipofectamin 2000. These methods are well Icnown in the ait and e.g. described by Ausbel et al. (eds.), 1996, Current Protocols in Molecular Biology, John Wiley & Sons, New York, USA. The cultivation of mammalian cells are conducted according to established methods, e.g. as disclosed in (Animal Cell Biotechnology, Methods and Protocols, Edited by Nigel Jenldns, 1999, Human Press Inc, Totowa, New Jersey, USA and Harrison MA and Rae IF, General Techniques of Cell Culture, Cambridge University Press 1997). In the production methods of the present invention, tlie cells are cultivated in a nutrient medium suitable for production of the polypeptide using methods Icnown in the art. For example, the cell may be cultivated by shake flask cultivation, small-scale or large-scale fermentation (including continuous, batch, fed-batch, or solid state fermentations) in laboratory or industrial fermenters performed in a suitable medium and under conditions allo^^'ing the polypeptide to be expressed and/or isolated. The culti\'ation takes place in a suitable nutrient medium comprising carbon and nitrogen sources and inorganic salts, using procedures known SO ' in the art. Suitable media are available from commercial suppliers or may be prepared according to published compositions (e.g., in catalogues of the American Type Culture Collection). If the polypeptide is secreted into the nutrient medium, the polypeptide can be recovered directly from the medium. If the polypeptide is not secreted, it can be recovered from :;ell lysates. The resulting polypeptide may be recovered by methods known in the art. For sxample, the polypeptide may be recovered from the nutrient medium by conventional procedures including, but not limited to, centrifugation, filtration, extraction, spray drying, evaporation, or precipitation. The polypeptides may be purified by a variety of procedures known in the art including, but not limited to, chromatography (e.g., ion exchange, affinity, hydrophobic, :hromatofocusing, and size exclusion), electrophoretic procedures (e.g., preparative isoelectric focusing), differential solubility (e.g., ammonium sulfate precipitation), SDS-PAGE, or sxtraction (see, e.g., Protein Purification, J.-C. Janson and Lars Ryden, editors, VCH Publishers, New York, 1989). Single chain FVII can be purified and activated to two-chain FVUa by a number of methods as described in the literature (Broze and Majerus, 1980, J. Biol. Chem. 255:1242-47 and Hedner and Kisiel, 1983, J.Clin.Invest. 71:1836-41). Another method whereby single chain FVn can bfi purified is by incorporation of Zn ions during purification as described in US 5,700,914. In a preferred embodiment the polypeptide is purified as a single chain FVH, which further is PEGylated, The PEGylated FVH single chain polypeptide is activated by either use of an immobilized enzyme (e.g. factors Ea, IXa, Xa and XHa) or by autoactivation using a positively charged ion exchange matrix or the like. It is advantageous to first puri:fy FVH in its single chain form, then PEGylate (if desired) and last activate by one of the methods described above or by autoactivation as described by Pedersen et al, 1989, Biochemistry 28: 9331-36. The advantage of carrying out PBGylation before activation is that PEGylation of the new aminoterminal formed by cleavage of R152-I153 is avoided, PEGylation of this new amino terminal would render the molecule inactive since the formation of a hydrogen bond between D242 and the amino terminal of 1153 is necessary for activity. Phannaceutical composition of the invention and its use Si In a further aspect, the present invention relates to a composition, in particular to a pharmaceutical composition, comprising a polypeptide or conjugate (including an inactive conjugate as described further above) of the invention and a pharmaceutically acceptable carrier or excipient. The conjugate, the polypeptide or the pharmaceutical composition according to the invention may be used as a medicament. Preferably, the pol3'peptide or the (active) conjugate may be used for the manufacture of a medicament for the treatment or prophylaxis of a FVIIa/TF-related disease or disorder in a mammal. For example, the polypeptide or the (active) conjugate may be used for the manufacture of a medicament for the treatment or prophylaxis of diseases wherein increased clot formation is desirable, such as treatment of patients having diseases resulting in inadequate blood coagulation in response to damage to blood vessels. In particular, the polypeptide or the (active) conjugate may be used for the manufacture of a medicament for the treatment of hemophiliacs, hemophiliacs with inhibitors to FVm and HX, patients with thrombocytopenia, patients with thromboc)^opathies, such as Glanzmann's thrombastenia platelet release defects and storage pool defects, patients with von Willebrand's disease, patients with liver diseases, or otherwise healthy people with severe bleeding problems, e.g. due to trauma or major surgery, who have developed inhibitors to FVTla, bleeding disorders such as hemophiliacs and other typically associated with severe tissue damages. Analogously, the inactive conjugate of the invention may be used for the manufacture of a medicament for the treatment or prophylaxis of a FVIIa/TF-related disease or disorder in a mammal. For example, the inacti^'e conjugate of the invention may be used for the manufacture of a medicament for the treatment or prophylaxis of diseases where decreased clot formation is desirable, such as prophylaxis or treatment of patients being in hypercoagulable states, such as patients with sepsis, deep-vein thrombosis, patients in risk of myocardial infections or thrombotic stroke, pulmonary embolism, patients with acute coronary syndromes (myocardial infarction and unstable angina pectoris), patients undergoing coronary cardiac, prevention of cardiac events and restonosis for patients receiving angioplasty, patients with peripheral vascular diseases. The inactive conjugate of the invention may also be used for the manufacture of a medicament for the treatment of respirator}' diseases, tumor growth and metastasis. In another aspect, the polx'peptide, the (active) conjugate or the pharmaceutical composition comprising the (active) conjugate of the invention may be used in a method for treating a mammal having a F\TIa/TF-relaied disease or disorder (such as one or more of the sx > diseases or disorders mentioned above), comprising administering to a mammal in need thereof an effective amount of such a polypeptide, conjugate or composition. Analogously, the inactive conjugate or the pharmaceutical composition comprising the inactive conjugate of the invention may be used in a method for treating a mammal having a FVna/TF-related disease or disorder (such as one or more of the diseases or disorders mentioned above), comprising administering to a mammal in need thereof an effective amount of such an inactivated conjugate or composition. The polypeptides or conjugates of the invention is administered to patients in a therapeutically effective dose, normally one approximately paralleling that employed in therapy with rFVn such as NovoSeven®, or at higher dosage. By "therapeutically effective dose" herein is meant a dose that is sufficient to produce the desired effects in relation to the condition for which it is administered. The exact dose will depend on the circumstances, and will be ascertainable by one skilled in the art using known techniques. Normally, the dose should be capable of preventing or lessening the severity or spread of the condition or indication being treated. It will be apparent to those of skill in the art that an effective amount of a polypeptide, conjugate or composition of the invention depends, inter alia, upon the disease, the dose, the administration schedule, whether the polypeptide or conjugate or composition is administered alone or in conjunction with other therapeutic agents, the plasma half-life of the compositions, and '"hf. crp.nftrfll hp.alth nf fhft natient.. Preferably, the DolvneDtide. coniusate. or comoosition of the invention is administered in an effective dose, in particular a dose which is sufficient to normalize the coagulation disorder. The polypeptide or conjugate of the invention is preferably administered in a composition including a pharmaceutically acceptable carrier or excipient. 'Tharmaceutically acceptable" means a carrier or excipient that does not cause any untoward effects in patients to whom it is administered. Such pharmaceutically acceptable carriers and excipients are well known in the art (see, for example, Remington's Pharmaceutical Sciences, 18th edition, A. R. Gennaro, Ed., Mack Publishing Company [1990]; Pharmaceutical Formulation Development of Peptides and Proteins, S. Frokjaer and L. Hovgaard, Eds., Taylor & Francis [2000]; and Handbook of Pharmaceutical Excipients, 3rd edition, A. Kibbe, Ed., Pharmaceutical Press [2000]). The polypeptide or conjugate of the invention can be fomiulated into pharmaceutical compositions by well-knoviu methods. Suitable formulations are described by Remington's Pharmaceutical Sciences by E.W. Martin (Mark Publ. Co., 16th Ed., 1980). 53 The polypeptide or conjugate of the invention can be used "as is" and/or in a salt form thereof. Suitable salts include, but are not limited to, salts with alkali metals or alkaline earth metals, such as sodium, potassium, calcium and magnesium, as well as e.g. zinc salts. These salts or complexes may by present as a crystalline and/or amorphous structure. The pharmaceutical composition of the invention may be administered alone or in conjunction with other therapeutic agents. These agents may be incorporated as part of the same pharmaceutical composition or may be administered separately from the polypeptide or conjugate of the invention, either concurrently or in accordance with another treatment schedule. In addition, the pol}'peptide, conjugate or pharmaceutical composition of the invention may be used as an adjuvant to other therapies. A "patient" for tlie purposes of the present invention includes both humans and other mammals. Thus the methods are applicable to both human therapy and veterinary applications. The pharmaceutical composition of the polypeptide or conjugate of the invention may be formulated in a variety of forms, e.g. as a liquid, gel, lyophilized, or as a compressed solid. The preferred form will depend upon the particular indication being treated and will be apparent to one sldlled in the art. In particular, the pharmaceutical composition of the polypeptide or conjugate of the invention may be formulated in lyophilised or stable soluble form. The polypeptide or the conjugate may be lyophilised by a variety of procedures Icnown in the art. A polypeptide or the conjugate may be a stable soluble form by the removal or shielding of proteolytic degradation sites. The advantage of obtaining a stable soluble preparation Ues in easier handling for the patient and, in the case of emergencies, quicker action, which potentially can become life saving. The preferred form will depend upon the particular indication being treated and will be apparent to one of skill in the art. The administration of the formulations of the present invention can be performed in a variety of ways, including, but not limited to, orally, subcutaneously, intravenously, intracerebral!y, intranasally, transdermally, intraperitoneally, intramuscularly, intrapulmonaiy, vaginally, rectally, intraocularly, or in any other acceptable manner. The formulations can be administered continuously by infusion, although bolus injection is acceptable, using techniques well Icnown in the art, such as pumps or implantation. In some instances the formulations may be directly applied as a solution or spray. Parentals ■Sjf A preferred example of a pharmaceutical composition is a solution designed for parenteral administration. Although in many cases pharmaceutical solution formulations are provided in liquid form, appropriate for immediate use, such parenteral formulations may also be provided in frozen or in lyophilized form. In the former case, the composition must be thawed prior to use. The latter form is often used to enhance the stability of the active compound contained in the composition under a wider variety of storage conditions, as it is recognized by those skilled in the art that lyophilized preparations are generally more stable than their liquid counterparts. Such lyophilized preparations are reconstituted prior to use by the addition of one or more suitable pharmaceutically acceptable diluents such as sterile water for injection or sterile physiological saline solution. In case of parenterals, they are prepared for storage as lyophilized formulations or aqueous solutions by niixing, as appropriate, the polypeptide having the desired degree of purity with one or more pharmaceutically acceptable carriers, excipients or stabilizers typically employed in the art (all of which are termed "excipients"), for example buffering agents, stabilizing agents, preservatives, isotonifiers, non-ionic surfactants or detergents, antioxidants and/or other miscellaneous additives. Buffering agents help to maintain the pH in the range which approximates physiological conditions. They are typically present at a concentration ranging from about 2 mM to about 50 mM Suitable buffering agents for use with the present invention include both organic and inorganic acids and salts thereof such as citrate buffers (e.g., monosodium citrate-disodium citrate mixture, citric acid-trisodium citrate mixture, citric acid-monosodium citrate mixture, etc.), succinate buffers (e.g., succinic acid-monosodium succinate mixture, succinic acid-sodium hydroxide mixture, succinic acid-disodium succinate mixture, etc.), tartrate buffers (e.g., tartaric acid-sodium tartrate mixture, tartaric acid-potassium tartrate mixture, tartaric acid-sodium hydroxide mixture, etc.), fumarate buffers (e.g., fumaric acid-monosodium fumarate mixture, fumaric acid-disodium fumarate mixture, monosodium fumarate-disodium fumarate mixture, etc.), gluconate buffers (e.g., gluconic acid-sodium glyconate mixture, gluconic acid-sodium hydroxide mixture, gluconic acid-potassium glyuconate mixture, etc.), oxalate buffer (e.g., oxalic acid-sodium oxalate mixture, oxalic acid-sodium hydroxide mixture, oxalic acid-potassium oxalate mixture, etc.), lactate buffers (e.g., lactic acid-sodixmi lactate mixture, lactic • acid-sodium hydroxide mixture, lactic acid-potassium lactate mixture, etc.) and acetate buffers (e.g., acetic acid-sodium acetate mixture, acetic acid-sodium hydroxide mixture, etc.). Additional possibilities are phosphate buffers, histidine buffers and trimethylamine salts such as Tris. ss- Stabilizers refer to a broad categOT}' of excipients which can range in function from a bulldng agent to an additive which solubilizes the therapeutic agent or helps to prevent denaturation or adherence to the container wall. Typical stabilizers can be polyhydric sugar alcohols (enumerated above); amino acids such as arginine, lysine, glycine, glutamine, asparagine, histidine, alanine, omithine, L-leucine, 2-phenylalanine, glutamic acid, threonine, etc., organic sugars or sugar alcohols, such as lactose, trehalose, stachyose, mannitol, sorbitol, xylitol, ribitol, myoinisitol, galactitol, glycerol and the like, including cyclitols such as inositol; polyethylene glycol; amino acid polymers; sulfur-containing reducing agents, such as urea, glutathione, thioctic acid, sodium thioglycolate, thioglycerol, a-monothioglycerol and sodium thiosulfate; low molecular weight polypeptides (i.e. Preservatives are added to retard microbial growth, and are typically added in amounts of about 0.2%-1% (w/v). Suitable preservatives for use with the present invention include phenol, benzyl alcohol, meta-cresol, methyl paraben, propyl paraben, octadecyldimethylbenzyl ammonium cliloride, benzalkonium halides (e.g. benzallconium chloride, bromide or iodide), hexamethonium chloride, allcyl parabens such as methyl or propyl paraben, catechol, resorcinol, cyclohexanol and 3-pentanol. Isotonicifiers are added to ensure isotonicity of hquid compositions and include polyhydric sugar alcohols, preferably tiihydric or higher sugar alcohols, such as glycerin, erythritol, arabitol, xyUtol, sorbitol and mannitol. Polyhydric alcohols can be present in an amount between 0.1% and 25% by weight, typically 1% to 5%, taldng into account the relative amounts of the other ingredients. Non-ionic surfactants or detergents (also known as "wetting agents") may be present to help solubilizing the therapeutic agent as well as to protect the therapeutic polypeptide against agitation-induced aggregation, which also permits the formulation to be exposed to shear surface stress without causing denaturation of the polj'peptide. Suitable non-ionic surfactants include polysorbates (20, 80, etc.), polyoxamers (184, 188 etc.), Pluronic® polyols, polyoxyethylene sorbitan monoethers (Tween®-20, Tween®-80, etc.). SG Aaoitional miscellaneous excipients include bullang agents or tillers (e.g. starch), chelating agents (e.g. EDTA), antioxidants (e.g., ascorbic acid, methionine, vitamin E) and cosolvents. The active ingredient may also be entrapped in microcapsules prepared, for exainple, by coascervation techniques or by interfacial polymerization, for example hydroxymethylcellulose, gelatin or poly-(methylmethacylate) microcapsules, in colloidal dmg delivery systems (for example liposomes, albumin microspheres, microemulsions, nano-particles and nanocapsules) or in macroemulsions. Such techniques are disclosed in Remington's Pharmaceutical Sciences, supra. Parenteral formulations to be used for in vivo administration must be sterile. This is readily accomphshed, for example, by filtration through sterile filtration membranes. Sustained release preparations Examples of sustained-release preparations include semi-permeable matrices of solid hydrophobic polymers containing the polypeptide or conjugate, the matrices having a suitable form such as a film or microcapsules. Examples of sustained-release matrices include polyesters, hydrogels (for example, poly(2-hydroxyethyl-methacrylate) or poly(vinylalcohol)), polylactides, copolymers of L-glutamic acid and ethyl-L-glutamate, non-degradable ethylene-vinyl acetate, degradable lactic acid-glycolic acid copolymers such as the ProLease® technology or Lupron Depot® (injectable microspheres composed of lactic acid-glycolic acid copolymer and leuproUde acetate), and poly-D-(-)-3-hydroxybutyric acid. While polymers such as ethylene-vinyl acetate and lactic acid-glycolic acid enable release of molecules for long periods such as up to or over 100 days, certain hydrogels release proteins for shorter time periods. When encapsulated polypeptides remain in the body for a long time, they may denature or aggregate as a result of exposure to moisture at 37°C, resulting in a loss of biological activity and possible changes in immunogenicity. Rational strategies can be devised for stabilization depending on the mechanism involved. For example, if the aggregation mechanism is discovered to be intermolecular S-S bond formation through thio-disulfide interchange, stabilization may be achieved by modifying sulfhydryl residues, lyophilizing from acidic solutions, controlling moisture content, using appropriate additives, and developing specific polymer matrix compositions. The invention is further described in the following non-limiting examples. 5Y SEQ ID N0:1 shows the hFVH protein (including y-carboxylated residues) SEQ ID NO:2 shows the cDNA sequence encoding the hFVn protein. SEQ ID N0:3 shows the hFVII protein (without y-carboxylated residues) SEQ ID N0:4 shows an expression cassette for expression of FVH in mammalian ceils. SEQ ID N0:5 shows the CBProFprl74 primer. SEQ ID N0:6 sho\^'s the CBProFprl75 primer. SEQ ID N0:7 sho^^'s the CBProFpr216 primer. SEQ ID-N0:8 shows tlie CBProFpr229 primer. SEQ ID N0:9 shows the CBProFpr221 primer. SEQ ID NO: 10 shows the CBProFpr228 primer. SEQ ID NO: 11 shows the CBProFpr226 primer. MATERIALS AND ISIETHODS Methods used to determine the amino acids to be modified Accessible Surface Area (ASA) The computer program Access (B. Lee and F.M.Richards, J. Mol.Biol. 55: 379-400 (1971)) version 2 (© 1983 Yale University) are used to compute the accessible surface area (ASA) of the individual atoms in the structure. This method typically uses a probe-size of 1.4A and defines the Accessible Surface Area (ASA) as the area formed by the center of the probe. Prior to this calculation all water molecules and all hydrogen atoms should be removed from the coordinate set, as should other atoms not directly related to the protein. Fractional ASA of side chain The fractional ASA of the side chain atoms is computed by division of the sum of the ASA of the atoms in the side chain v\'ith a value representing the ASA of the side chain atoms of that residue tj'pe in an extended Ala-x-Ala tripeptide (See Hubbard, Campbell & Thornton (1991) J.Mol.Biol.220,507-530). For this example the CA atom is regarded as a part of the side chain of Glycine residues but not for the remaining residues. The following table is used as standard 100% ASA for the side chain: 5 f~> Ala 69.23 A^ Leu 140.76 A^ Arg 200.35 A^ Lys 162.50 A^ Asn 106.25 A^ Met 156.08 K^ Asp 102.06 A' Phe 163.90 A^ Cys 96.69 A^ Pro 119.65 A' Gin 140.58 A^ Ser 78.16 A^ Glu 134.61 A^ Thr 101.67 A^ Gly 32.28 A^ Trp 210.89 A^ His 147.00 A^ Tyr 176.61 A^ He 137.91 A^ Val 114.14 A^ Residues not detected in the structure are defined as having 100% exposure as they are thought to reside in flexible regions. The gamma-carboxy glutamic acids at positions 6, 7,14, 16,19, 20,25, 26, 29 and 35 are all defined as being 100% exposed. Determining distances betv[>een atonis The distance between atoms is most easily determined using molecular graphics software e.g. Insight!!® v. 98.0, MS! INC. Catalytic site region The catalytic site region is defined as any residues having at least one atom within 10 A of any atom in the catalytic triad (residues H193, D242, S344). Detertnination of tissue factor binding site The receptor-binding site is defined as comprising of all residues having their accessible surface area changed upon receptor binding. This is determined by at least two ASA calculations; one on the isolated ligand(s) in the ligand(s)/receptor(s) complex and one on the complete ligand(s)/receptor(s) complex. Methods for testing ¥VQ. and FVIIa properties Measurement offimctional in vivo lialf-life S^ Measurement oi in vivo Dioiogicai nair-iiie can oe carnea out m a numoer oi ways as described in the literature. An example of an assay for the measurement of i7i vivo half-life of rPs^a or variants thereof is described in EDA reference number 96-0597. Briefly, FVTE clotting activities are measured in plasma drawn prior to and during a 24-hour period after administration of the conjugate, polypeptide or composition. The median apparent volume of distribution at steady state is measured and the median clearance determined. Measurement of reduced sensitivity to proteolytic degradation A composition containing the conjugate (100-750 [xg/ml, preferably 600 [i-g/ml), 1.5 mg Ca'Vml (as calcium chloride), mannitol (30 mg/ml), polysorbate 80 (0.1 mg/ml), sodium chloride (3 mg/ml) and glycyl-glycine buffer (1.3 mg/ml, pH 5.5) is prepared. A similar composition containing wild-tj'pe rFVIIa is prepared. The initial clotting activity or, alternatively, the initial amidolytic activity is then determined as described in the section entitled "Methodfor measuring the clotting activity" or as described in the section entitled "Method of measuring low levels of catalytic activity" or "Method ofmeaswing the catalytic activity". The compositions are then incubated at 37°C until the composition containing the wild-type rF\TI has lost at least 25%, preferably at least 50%, of its initial clotting or amidolytic acitivity. The clotting or amidolytic activity of the composition containing the conjugate of the invention is then measured. The reduced sensitivitj' to proteoljdc degradation of the conjugate of the invention as compared to wild-tj'pe rFVIIa is then expressed in percentage. Alternative methods for measwing reduced sensitivity to proteolytic degradation Proteolytic degradation can be measured using the assay described in US 5,580,560, Example 5, where proteolysis is autoproteolysis. Furthermore, reduced proteolysis can be tested in an in vivo model using radiolabelled samples and comparing proteolysis of wild type and conjugates by withdrawing blood samples and subjecting these to SDS-PAGE and autoradiography. Irrespectively of the assay used for determining proteolytic degradation, "reduced proteolytic degradation" is intended to mean a measurable reduction in cleavage compared to that obtained by non-conjugated wild t^pe F\T[a as measured by gel scanning of Coomassie 60 stained HUH-fAUt. gels, tiFLL: or as measured by conserved catalytic activity m comparison to wild type using the chromogenic assay described below. Determination of the molecular weight ofrFVUand conjugates thereof The molecular weight of conjugated or unconjugated rFVn or conjugates thereof is determined by either SDS-PAGE, gel filtration, Westem Blots, matrix assisted laser desorption mass spectrometry or equilibrium centrifugation, e.g. SDS-PAGE according to Laemmli, U.K., Nature Vol 227 (1970), pp. 680-85. Method of measuring low levels of catalytic activity The amidolytic activity in dilute samples of FVB/FVlIa and fermentation liquid/conditioned medium can be determined using CO ASET® FVH (Chromogenix, Art. No 82 19 00). The amidolytic activity is determined in accordance with the manufacturer's instractions. Briefly, FX is present in surplus and converted to FXa by FVIIa at 37°C. The generated FXa then hydrolyses the chromogenic substrate S2765 (N-a-Cbo-D-Arg-Gly-Arg-pNA) resulting in liberation of the chromophoric molecule, para-nitro-anilHn (pNA) absorbing light at wavelength W5 nm. The reaction is stopped by addition of acetic acid. The amount of FVE/FVlIa in the sample is determined by comparison to a standard curve prepared from FVlIa (ranging from 125 pg/ml to 1 ng/ml in assay buffer). Method of measuring the catalytic activity The ability of the conjugates to cleave small peptide substrates can be measured using the chromogenic substrate S-2288 (D-He-Pro-Arg-p-nitroanilide). Recombinant FVlIa is diluted in 0.1 M Tris, 0.1 M NaCl, 5 mM CaCh, pH 8.3 containing 0.1% BSA. The reaction is started by addition of the S-2288 substrate to 1 mM and the absorption at 405 lun is measured after incubation for 30 min. at 37°C. Method of measuring the clotting activity FSQIa activity is measured using a standard one-stage clotdng assay essentially as described in W092/156S6. Briefly, the sample to be tested is diluted in 50 mM Tris (pH 7.5), D.1% BSA and 100 ^1 is incubated with 100 pi of FVn deficient plasma and 200 yil of thromboplastin C containing 10 mM Ca"*^. Clotting times are measured and compared to a standard curve using a pool of citrated normal human plasma in serial dilution. Method of measuring the anticoagulant activit)' The anticoagulant activit}' of an inactive FVH or FVHa conjugate can be measured using the one-stage clotting assay described above (MetJiod of measuring the clotting activity) where the inactive conjugate competes with wild-type FVn for a limited amount of relipidated tissue factor. The assay is performed essentially as described in WO 92/15686, example HI, which is hereby incorporated as reference. The ability of the inactive conjugate to prolong the clotting time of wild-type FVII is recorded and taken as a measure of anticoagulant activity. EXAMPLES Example 1 The X-ray structure of hFVIIa in complex with soluble tissue factor by Banner et.al., J Mol Biol, 1996; 285:2089 is used for this example. It is noted that the numbering of residues in the reference does not follow the sequence. Here we have used the sequential numbering according to SEQ ID N0:1. The gamma-carboxy glutamic acids at positi5ns 6, 7,14, 16,19, 20, 25, 26,29 and 35 are all here named GLU (three letter abbreviation) or E (one letter abbreviation). Residues 143-152 ai-e not present in the stmcture. Swface exposure Performing fractional ASA calculations on FVII fragments alone combined with the definition of accessibilities of non standard and/or missing residues described in the methods resulted in the following residues having more than 25% of their side chain exposed to the surface: Al, N2, A3, F4, L5, E6, E7, L8, R9, PIG, S12, L13, E14, E16, K18, E19, E20, Q21, S23, F24, E25, E26, R28, E29, F31, K32, D33, A34, E35, R36, K38, L39, W41,142, S43, 545, 047, D48, Q49, A51, S52, S53, Q56, 058, S60, K62, D63, Q64, L65, Q66, S67,169, F71, L73, P74, A75, E77, G78, R79, E82, T83, H84, K85, D86, D87, QS8, L89,190, V92, N93, E94, 097, E99, S103, D104, H105, T106, G107, T108, K109, Sill, R113, El 16, G117, S119, L120, L121, A122, D123, G124, V125, S126, T128, P129, T130, V131, E132,1140, L141, E142, K143, R144, N145, A146, S147, K148, P149, Q150, 0151, R152, 0155, K157, V158, F160, K161, E163, L171, N173, G174, A175, N184, T185,1186, H193, K197, K199, N200, R202, N203,1205, S214, E215, H216, D217, 0218, D219, S222, R224, S232, T233, V235, P236, G237, T238, T239, N240, H249, Q250, P251, V253, T255, D256, E265, R266, T267, E270, R27L F275, V276, R277, F278, L280, L287, L288, D289, R290, 0291, A292, T293, L295, G3b E296, N301, M306, T30/, ^Mn, ujuy, L^iii, KI^IA V-3-I-^> is-ju, js^io, V^JI /, ^JJio, L^JJ.^, S320, P321, N322, T324, E325, Y326, Y332, S333, D334, S336, K337, K341, G342, H351, R353, G354, Q366, G367, T370, V371, G372, R379, E385, Q388, K389, R392, S393, E394, P395, R396, P397, G398, V399, L400, L401, R402, P404 and P406. i The following residues had more than 50% of their side chain exposed to the surface: Al, A3, F4, L5, E6, E7, L8, R9, PIO, E14, E16, K18, E19, E20, Q21, S23, E25, E26, E29, K32, A34, E35, R36, K38, L39,142, S43, G47, D48, A51, S52, S53, Q56, G58, S60, K62, L65, Q66, S67,169, F71, L73, P74, A75, E77, G78, R79, E82, H84, K85, D86, D87, Q88, L89,190, V92, N93, E94, G97, T106, G107, T108, K109, Sill, E116, S119, L121, A122, D123, G124, V131, i E132, L141, E142, K143, R144, N145, A146, S147, K148, P149, Q150, G151, R152, G155, K157. P160, N173, G174, A175, K197, K199, N200, R202, S214. E215, H216, G218, R224, V235, P236, G237, T238, H249, Q250, V253, D256, T267, F275, R277, F278, L288, D289, R290, G291, A292, T293, L295, N301, M306, Q308, D309, L311, Q312, Q313, R315, K316, G318, D319, N322, E325, D334, K341, G354, G367, V371, E385, K389, R392, E394, R396, I P397, G398, R402, P404 and P406. Tissue factor binding site Performing ASA calculations the following residues in human FVH change their ASA in the complex. These residues were defined as constituting the receptor binding site: L13, K18, a F31, E35, R36. L39, F40,142, S43, S60, K62, D63, Q64, L65,169, C70, F71, C72, L73, P74, F76, E77, G78, R79, E82, K85, Q88,190, V92, N93, E94, R271, A274, F275, V276, R277, F278, R304, L305, M306, T307, Q308, D309, Q312, Q313, E325 and R379. Active site region i The active site region is defined as any residue having at least one atom within a distance of 10 A from any atom in the catalytic triad (residues H193, D242, S344): 1153, Q167, V168, L169, L170, L171, Q176, L177, C178, G179, G180, T181, V188, V189, S190, A191, A192, H193, C194, F195, D196, K197,1198, W201, V228.1229,1230, P231, S232, T233, Y234, V235, P236, G237, T238, T239, N240, H241, D242,1243, A244, L245, L246, V281, (f S282, G283, W284, G285, Q286, T293, T324, E325, Y326, M327, F328, D338, S339, C340, K341, G342, D343, S344, G345, G346, P347, H348, L358, T359, G360,1361, V362, S363, W364, G365, C368, V376, Y377, T378, R379, V380, Q382, Y383, W386, L387, L400 and F405. 63 Tlie ridge of the active site binding cleft The ridge of the active site binding cleft region was defined by visual inspection of the FVna structure lFAK.pdb as: N173, A175, K199, N200, N203, D289, R290, G291, A292, P321andT370. Example 2 Design of an expression cassette for expression of human blood coagulation factor VII in nuanmalian cells The DNA sequence shown in SEQ ID N0:2, encompassing the short form of the full length cDNA encoding human blood coagulation factor VH with its native short signal peptide (Hagen et al., 1986. PNAS 83:2412), was synthesized in order to facilitate high expression in mammalian cells. First the ATG start codon context was modified according to the Kozalc consensus sequence (Kozak, M.JM0I Biol 1987 Aug 20;196(4):947-50), so that there is a perfect match to the consensus sequence upstream of the ATG start codon. Secondly the open "reading frame of the native human blood coagulation factor cDNA was modified by maldng a bias in the codon usage towards the codons frequently used in highly expressed human genes. Further, two translational stop codons was inserted at the end of the open reading frame in order to facihtate efficient translational stop. The fully synthetic and expression optimized human FVn gene was assembled from 70-mer DNA oligonucleotides and finally amplified using end primers inserting BamlE and HiitdEL sites at the 5' and 3' ends respectively using standard PCR techniques, which resulted in the following sequence (SEQ ID NO:4): ggatcccgccaccatggtcagccaggccctccgcctcctgtgcctgctcctggggctgcagggctgcctggctgccgtcttcgtcaccca ggaggaagcccatggcgtcctgcatcgccggcgccgggccaatgcctttctggaagagctccgccctggctccctggaacgcgaatgc aaagaggaacagtgcagctttgaggaagcccgggagattttcaaagacgctgagcggaccaaactgttttggattagctatagcgatggc gatcagtgcgcctccagcccttgccagaacgggggctcctgcaaagaccagctgcagagctatatctgcttctgcctgcctgcctttgagg ggcgcaattgcgaaacccataaggatgaccagctgatttgcgtcaacgaaaacgggggctgcgagcagtactgcagcgatcacacggg cacgaagcggagctgccgctgccacgaaggctatagcctcctggctgacggggtgtcctgcacgcccacggtggaatacccttgcggg aagattcccattctagaaaagcggaacgctagcaaaccccagggccggatcgtcggcgggaaggtctgccctaagggggagtgcccct ggcaggtcctgctcctggtcaacggggcccagctgtgcggcgggaccctcatcaataccatttgggtcgtgtccgccgctcactgcttcg ataagattaagaattggcggaacctcatcgctgtgctcggcgaacacgatctgtccgagcatgacggggacgaacagtcccgccgggtg gctcaggtcatcattccctccacctatgtgcctggcacgaccaatcacgatatcgctctgctccgcctccaccagcccgtcgtgctcaccga tcacgtcgtgcctctgtgcctgcctgagcggacctttagcgaacgcacgctggctttcgtccgctttagcctcgtgtccggctggggccag GJf ctgctcgaccggggcgctaccgctctcgagctgatggtgctcaacgtcccccggctgatgacccaggactgcctgcagcagtcccgcaa agtgggggactcccccaatatcacggagtatatgttttgcgctggctatagcgatggctccaaggatagctgcaagggggactccggcg ggccccatgccacgcactatcgcgggacctggtacctcaccgggatcgtcagctggggccagggctgcgccacggtggggcactttg gcgtctacacgcgcgtcagccagtacattgagtggctgcagaagctcatgcggagcgaaccccggcccggggtgctcctgcgggccc f ctttcccttgataaaagctt A vector for the cloning of the generated PCR product encompassing the expression cassette for native human blood coagulation factor Vn was prepared by cloning the intron from pCINeo (Promega). The synthetic intron from pCI-Neo was amplified using standard PCR 5 conditions as described above and the primers; CBProFprl74: 5'- AGCTGGCTAGCCACTGGGCAGGTAAGTATCA -3' and CBPwFprl75: 5'- TGGCGGGATCCTTAAGAGCTGTAATTGAACT -3' ? resulting in a 332 bp PCR fragment. The fragment was cut with Nliel and BairiHl before cloning into pCDNA3.1/HygR (obtained from In Vitro Gen) resulting in PF#34. The expression cassette for native human blood coagulation factor Vn was cloned between the BamEI and Hindlll sites of PF#34, resulting in plasmid PF#226. i Example 3 Construction of expression cassettes encoding variant forms of human blood coagulation factor VII that are having an additional in vivo glycosylation site. Sequence overhang extension (SOE) PCR was used for generating constructs having t variant human blood coagulation factor open reading frames with substituted codons. In the SOE-PCR both the N-tenrdnal part and the C-terminal part of the FVn open reading frame was first amplified in individual primary PCRs. For example, in order to change the codons for R315 and V317 to the codons forN315 and T317 the following primers were used pair vice for the primary PCR's: i CBProFpr2l6: 5'-CTTAAGGATCCCGCCACCATGGTCAGCCAG-3' and CBPi-oFpr229: 5'-GGAGTCCCCGGTTTTGTTGGACTGCTGC-3', and CBProFpr221: 5'-ACTTAAGCTTTTATCAAGGGA-3' and (.^ , CBProFpr228: 5'-GCAGCAGTCCAACAAAACCGGGGACTCC-3'. The primary PCR products were then combined and the terminal primers (CBProFpr216 and CBProFpr221) added resulting in the secondary full-length product i encoding the desired R315N + V317T FVU variant. The secondary PCR product was trimmed with BamHI and HindlH before cloning into the vector PF#34 between the BamBl and HindHl sites resulting in PF#249. Furthermore, in cases where the introduced mutation(s) were sufficiently close to a unique restriction endo-nuclease site in the expression plasmid variant genes were constructed I using construction procedure encompassing a single PCR step and a subsequent cloning. For instance, the substitution K143N+N145T was introduced by use of the PCR primers: CBProFpr226: 5'-CATTCTAGAAAACCGGACCGCTAGCAAACC-3' and CBProFpr221: 5'- ACTTAAGCTTTTATCAAGGGA-3' f i in a single PCR reaction. The PCR product was subsequently cloned using the restriction endo-nuclease sites Xbal and HindlR. Using the above strategy, the following glycosylation conjugates were prepared and ^ their amidolytic activities were tested as described in the section entitled Method of measuring low levels of catalytic acth'ity. Furthermore, some of the conjugates were subjected to the one-stage clotting assay described in the section entitled Method of measuring the clotting activity. The obtained results are compiled in the below table. +: Acitivty measurable; -: Activity not measurable; nd: not determined Example 4 f Glycosylation conjugate Amidolytic activity Clotting activity T106N + + K143N-FN145T + nd V253N + nd « R290N-KA292T - nd 029 IN - nd R315N+V317T + + K143N+N145T+R315N+V317T + nd Improvement of glycosylation site utility by introduction of another proximal (N-tenninally located) glycosylation site. In order to prevent autolysis of wild type human FVH a glycos3'iation site was introduced at position 315 by making the substitutions: R315N and V317T as described above, resulting in PF#249. Upon transfection of CHO Kl cells, using lipofactamine 2000, low transient expression levels were obtained. Assajing the twenty-four hour transient supernatant by the amidiolytic assay, COASET® FVn, indicated that the variant was active. After selection using 400 /xg/ml Hygromycin B a pool of stable clones was obtained. This pool expresses the R315N+V317T variant at approximately 0.2 /ig/ml allowing for Western blotting analysis for determination of the degree of usage of the introduced glycosylation site. A twenty-four hour supernatant from the stable pool was assayed by Western blotting and revealed partial usage of the introduced glycosylation site at position 315, approximately one-half of the fully processed secreted protein is glycosylated. However, if the native glycosylation site at position 145 is moved to position 143, by making the substitutions K143N+N145T, the introduced glycosylation site at position 315 is completely glycosylated as judged by Western blotting. Example 5 Expression ofFVII in HEK 293 cells The cell line HEK 293 (ATCC # CRL-1573) was seeded at 20% confluence in T-25 flasks using DMEM, high glucose 10% heat inactivated PCS (Gibco/BRL Cat # 10091), 5/ig/ml phylloquinone and allowed to grow until confluent. The confluent mono cell layer was transfected with 1, 2, 5,10 and 20 jig of the plasmid p226 described above using the Lipofectamine 2000 transfection agent (Life technologies) according to the manufactures instructions. After 24 hours post transfection a medium sample was drawn. The FVH concentration in the 24 hour transient expression experiments was on average 0.15 /ig/ml. Subsequently, selection medium containing 100 [ig/ml Hygromycin B was administered to the cells. After three weeks of selection, with renewal of the medium every 3rd or 4th day, the Hygromycin resistant cells were confluent in the flasks transfected with 1/ig of plasmid DNA and 2/x,g of plasmid DNA. The cells from each of the five flasks were harvested and pooled. The resulting stable pool of transfectants expressing native human blood coagulation factor Vn was frozen in liquid nitrogen according to standard procedures. 6^ Example 6 Generation ofHEK293 cells stably expressing FVII. A vial of HEK293 PF#226 transfectant pool was thawed and the cells seeded in a 75 crn^ tissue flask containing 15 ml of DMEM high glucose, 10% PCS, phylloquinone (5 fig/ml), 100 U/1 penicillin, 100/ig/l streptomycin, which was used for all subsequent experiments, and grown for 24 hours. The cells were harvested, diluted and plated in 96 well micro titer plates at a cell density of V2 ceU/well. After 12 days colonies of about 20-100 cells were present in the wells and those wells containing onl)' one colony were labeled. After a further two days of growth an additional 100 fil medium was added to all wells. Two days later the media in all wells containing only one colony were changed. The first colonies were transferred to 25 cm^ • tissue flaslcs culturing after 3 days and depending on the level of confluency the colonies were transferred to 25 cm^ tissue flasks culturing the next 11 days. When confluent in T-25 tissue flasks the medium was changed and clones allowed to secrete FVTI into the growth medium for 24 hours, after which the supematants were harvested and assayed for the presence of factor VU using the COASET FVII amidolytic assay. One clone, CIS, was found to express 29 /ig/ml Fvn. Example 7 Expression of FVII glycosylation variants Mnth no amidolytic activity capable of inhibiting the function ofF\'Tla The expression plasmids for expression of the active site ridge mutants R290N+A292T and G291N were constructed essentially as described in example 3. Using the amidiolytic assay COASET® FVII (see above) the ability of the two FVn glycosylation variants, R290N+A292T and G291N, to inhibit the activity of rFVUa was evaluated. The plasmids PF#250 encoding R290N+A292T and the plasmid PF#294 encoding G291N were transfected into near confluent serum grown HEK293 c&Us using Lipofectamin 2000. The transfected cells were incubated at 37°C witli 5% CO2 for three hours post transfection before the medium was changed to serum free medium (DMEM, ITS-A, ExCyte, Phylloquinone, P/S). Forty hours post transfection the conditioned medium was harvested and cleared by centrifugation for analysis. 6 Q A standard curve was made from rPVIIa: 0.0125 ng, 0.025 ng, 0.05 ng, 0.075 ng and 0.1 ng. Firfty fil aliquots of undiluted, 2-foId diluted, 5 fold diluted, 10-fold diluted and 50-fold diluted condition medium from either of the two inactive glycosylation variants R290N+A292T and G291N or conditioned medium from a mock transfection were spiked with 0.025 ng i rFVna. When assayed in the CO ASET FVn assay 0.025 ng of FVna corresponded to a signal of OD405 = 0.35 (first run) and OD405 = 0.26 (second run). The obtained results are compiled in the below table: Glycosylation conjugate Dilution OD405 ? Standard 0.35 0.26 Mock 50 0.38 0.19 25 0.31 0.16 i 10 0.21 0.15 5 0.22 0.14 1 0.08 0.07 R290N+A292T 50 0.23 _ D 25 0.12 - 10 0.07 - 5 0.04 - 1 0.04 - f G291N 50 „ 0.16 25 - 0.08 10 - 0.06 5 - 0.05 1 - 0.04 As it appears from the above data the glycosylation conjugates G291N and R290N+A292T inhibit the function rFVIIa. Example 8 3 Purification ofFVII and subsequent activation Purification of wild-type Factor VTl and conjugates thereof was performed at 4°C. The supernatant from cells expressing the conjugate (or wild-type FVE) was sterile filtered (0.22 l-im) and diluted 2 fold in cold milliQ water. EDTA was added to 5 mM, pH adjusted to 8.6 and D conductivity was below 10 mS/cm. The sample was applied onto a Q-sepharose Fast Flow resin (Pharmacia) equilibrated at 4°C in 10 mM Tris pH 8.6. After application of the sample the 6^ column was washed in 10 mM Tris (pH 8.6), 150 mM NaCl, until the absorption at 280 nm reached baseline levels. Then the column was equilibrated in 10 mM Tris (pH 8.6), 100 mM NaCl. Bound conjugate (or wild-type FVn) was eluted with 10 mM Tris (pH 8.6), 100 mM NaCl, 5 mM CaCla. Fractions enriched in conjugate (or wild-type FVH) was pooled and concentrated by dialysis or by using Vivaspin concentration units (Vivascience). Auto-activation of the conjugate (or wild-type FVH) was obtained by concentration and incubation of the eluted protein in 10 mM Tris (pH 7.8-8.6), 100 mM NaCl, 5 mM CaCh. Alternatively, the conjugate (or wild-type FVII) was activated at 37°C by Factor Xa coupled to CNBr-activated sepharose in 10 mM Tris (pH 7.4-8.0) 100 mM NaCl, 5 niM CaCl2. The conjugate (or wild-type FVHa) was buffer-exchanged into a solution containing 10 mM CaCb, 50 mM NaCl, 3% mannitol, 0.05% TweenSO, buffered at pH 5.6 and sterilfiltered before storage at -80°C. Example 9 N-tenninal PEGylation ofF\ni Factor VII was conjugated with methoxy polyethylene glycol (mPEG) having a molecular weight of about 5 IcDa using M-PEG-CHO (M-ALD-5000, obtained from Shearwater) in a buffer containing 10 mM sodium citrate, 20 mM CaC^, 100 mM NaCl, pH 5.5. M-PEG-CHO was present in 50-100-fold molar excess, and the protein concentration was 0.2-0.5 mg/ml. The reaction was carried out in 300-1500 [i,l batches at room temperature for 1 hour with agitation, and NaBHsCN was added to 500-1000 fold excess and incubation continued over night with agitation at room temperature. PEGylated FVII was buffer exchanged to buffer A (10 mM Tris pH 7.6) and applied at 4 °C on a mono Q column (Pharmacia) equilibrated in buffer A. Bound protein was eluted in a gradient from 0-100% B (10 mM Tris (pH 7.6), 500 mM NaCl) over 40 column volumes. Example 10 Pharmacokinetic studies in rats Both valid-type FVII and the conjugates of the invention are formulated in 1.3 mg/nol glycyl-glycin buffer pH 5.5 containing 1.5 mg/ml Carla, 30 mg/ml mannitol, 0.1 mg/ml polysorbat 80 and 3 mg/ml NaCl. For determination of in vivo half-life each of the preparations are administered to Sprague-Dawley rats as one intra venous bolus injection. The injections are 10 given slowly over about 10 seconds to reduce potential risk of heart failure due to high Ca ^* concentration. Blood samples are drawn from each of the nine anaesthetized rats, at suitable intervals, e.g. 1 minute, 15 minutes, 30 minutes, 45 minutes and 1 hour after injection. The blood samples are collected in 1 ml tubes containing 50 fall Citrate-phosphate-dextrose solution with adenine (Sigma #C4431) in order to prevent coagulation. Immediately after collection the samples are stored at about 0°C until centrifugation and subsequent collection of the citrate plasma supematants for assay. Samples are assayed by the one-stage clotting assay as described in the section Method of measuring the clotting activity and the half-lives are then calculated. WE CLAIM; 1. A polypeptide conjugate comprising a polypeptide having at least one sugar moiety covalently attached to an introduced in vivo N-glycosylation site of the polypeptide wherein the polypeptide part of said conjugate has an amino acid sequence that differs from that of wild-type FVII or FVIIa shown in SEQ ID NO:l in 1-15 amino acid residues and wherein said introduced in vivo N-glycosylation site is introduced by a substitution selected from the group consisting of F4S/T, PION, Q21N, W41N, S43N, A51N, G58N, L65N, G59S/T, E82S/r, N95S/r, G97S/T, YIOIN, D104N, T106N, K109N, G117N, G124N, S126N, T128N, A175S/r, G179N, I186S/r, V188N, R202S/r, I205S/r, D212N, E220N, I230N, P231N, P236N, G237N, V253N, E265N, T267N, E270N, R277N, L280N, G291N, P303S/T, L305N, Q312N, G318N, G331N, D334N, K337N, G342N, H348N, R353N, Y357N, I361N, V376N, R379N, M391N, and combinations thereof. 2. The polypeptide conjugate according to claim 1, wherein the substitution is selected from the group consisting of F4SAr, PION, Q21N, W41N, A51N, G58N, G59SAr, N95Sn, G97S/r, YIOIN, D104N, T106N, K109N, G117N, G124N, S126N, T128N, A175S/r, I186S/r, V188N, R202SA', I205S/T, D212N, E220N, V253N, E265N, T267N, E270N, L280N, G291N, P303S/T, G318N, G331N, D334N, K337N, R353N, Y357N, M391N, and combinations thereof. 3. The polypeptide conjugate according to claim 2, wherein the substitution is selected from the group consisting of T106N and V253N. 4. The polypeptide conjugate according to claim 1, wherein said in vivo N- glycosylation site is introduced in a position which does not form part of the tissue factor binding site, the active site region and the ridge of the active site binding cleft as defined herein. 5. The polypeptide conjugate according to any of claims 1-4, wherein two or more in vivo N-glycosylation sites have been introduced. 6. A nucleotide sequence encoding a polypeptide with an amino acid sequence as defined in any of claims 1-5. 7. A pharmaceutical composition comprising a polypeptide conjugate as defined in any of claims 1-5 and a pharmaceutical acceptable carrier or excipient.

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in-pct-2002-1373-che form-1.pdf

in-pct-2002-1373-che form-13.pdf

in-pct-2002-1373-che form-18.pdf

in-pct-2002-1373-che form-26.pdf

in-pct-2002-1373-che form-3.pdf

in-pct-2002-1373-che form-5.pdf

in-pct-2002-1373-che form-6.pdf

in-pct-2002-1373-che others.pdf

in-pct-2002-1373-che pct search report.pdf

in-pct-2002-1373-che pct.pdf

in-pct-2002-1373-che petition.pdf