PT1335938E - Apolipoprotein construct - Google Patents

Description

1

DESCRIPTION " CONSTRUCTIONS OF APOLIPOPROTEINS " The invention relates to an apolipoprotein construct, to an apolipoprotein construct usable as a medicament, to a nucleic acid sequence encoding the apolipoprotein construct, to a vector comprising the nucleic acid sequence, to a method for the production of A pharmaceutical composition comprising the preparation of apolipoprotein and the use of the apolipoprotein construct for the preparation of a pharmaceutical composition. BACKGROUND ART

In the present document, the term " Apo A " or " apolipoprotein A " will be used to denote any of the three apolipoproteins, apolipoprotein A, apolipoprotein A II or apolipoprotein A IV.

Cardiovascular diseases caused by atherosclerosis in vessels are the most frequent cause of death in the industrialized countries of the world. One of the pathogenic factors that causes atherosclerosis is the deposition of cholesterol in the walls of the vessels leading to the formation of plaques and eventually atherosclerosis and increasing the risk of infarction. Apolipoprotein A-1 (apo-A-1) is the major component of HDL (high-density lipoprotein) in plasma, which is negatively correlated with the presence of atherosclerosis. There is strong experimental evidence that this effect is caused by so-called reverse cholesterol transport from the peripheral tissues to the liver. There is also experimental evidence that this reverse cholesterol transport can be stimulated in mammals by injection with apo-A-1. Apolipoprotein A1 is rapidly removed from plasma, it is believed that much of Apo-a-1 is removed from the plasma by filtration in the kidneys without first being degraded (Braschi et al., 1999, J Lipid Res, 40: 522-532 , Braschi et al., 2000, Biochemistry, 39: 5441-5449; Glass et al 1983, J Biol Chem 258: 7161-7167). The short half-life of apolipoprotein A in plasma is an impediment to the use of the protein for the treatment of atherosclerosis.

US 5 876 968, WO 9312143 (SIRTORI ET AL.) Relates to substantially pure dimers of a variant of apo-A-1, designated apolipoprotein A-1-Milano. Medicaments containing the dimer may be used for the prevention of thromboses or may be used as prodrugs for the monomer. A specific feature of this particular variant of apo-A-i is its ability to form covalent dimers with itself. The authors speculated that the presence of Apo A-I-M could be responsible for a prolonged plasma half-life, but no conclusive data were presented.

In US 5 643 757 (SHA-IL ET AL.) There is disclosed a method for the production, with high yield, of pure, stable, mature and biologically active human apolipoprotein A-I. 3

In US 5 990 081 (AGELAND ET AL.) There is disclosed a method for the treatment of atherosclerosis or cardiovascular diseases by administering a therapeutically effective amount of apolipoprotein A or apolipoprotein E.

Human homologous dimers of apolipoprotein AI variants comprising a cysteine at position 151 are described in WO 96/37608 (RHONE-POULENC ROHRER ET AL.). The presence of the cysteine residue in the amino acid sequence allows the formation of dimers by means of disulfide bridges between the monomers. This reference further describes the corresponding amino acid sequences and the vectors comprising them, as well as the pharmaceutical compositions comprising the variants and the use thereof in gene therapy.

There are described in WO 90/12879 (Sirtori et al) and WO 94/13819 (Kabi Pharmacia) methods for the preparation of ApoA-1 and ApoA-1M in yeast and E. colif respectively. These documents also describe the use of ApoA-I and ApoA-IM as a medicament for the treatment of atherosclerosis and cardiovascular diseases.

In conclusion, the prior art is primarily concerned with the use of native ApoA-I or ApoA-IM monomer or ApoA-IM dimer as medicaments for the treatment of vascular diseases, despite the known drawbacks of these proteins quick debit). The prior art does not suggest modification of ApoA-I to provide constructs with increased ability to effect reverse cholesterol transport and / or a longer plasma half-life. It is thus an object of the present invention to provide such constructs of ApoA, which can be used for the treatment and / or prevention of cardiovascular diseases.

Short description

According to a first aspect, the invention relates to an apolipoprotein construct which can be used as a medicament having the general formula - apo A-X,

- wherein apo A is a component of apolipoprotein A selected from the group consisting of apolipoprotein AI, apolipoprotein AII, apolipoprotein IV, an analog or variant thereof, and X is a heterologous radical which comprises at least one compound selected from the group consisting of consisting of an amino acid, a peptide, a protein, a carbohydrate and an amino acid sequence, provided that if the construct consists of two identical native apolipoproteins, then these are serially linked. The invention provides a novel construction usable as a medicament. The prior art does not have an apolipoprotein construct as defined in the present invention for medical use. The apolipoprotein constructs of the present invention may be considered as HDL analogs because of their ability to form complexes with cholesterol and other lipids and their assistance in the transport of these compounds to the liver.

Throughout the invention, the apolipoprotein component or part of the construct is referred to as apo A or apolipoprotein. In the subsequent text and in the claims, the heterologous radical is referred to as component x of the construct. The apolipoprotein, or an analog or variant thereof, is covalently linked to the heterologous radical. Component X of the construct can be considered as a heterologous radical. In this context, a heterologous radical is a radical of any type that is not attached to the apolipoprotein, or an analog or variant or functional equivalent thereof, under native conditions. Thus, the heterologous radical may be a peptide or a protein or part of a peptide or protein of the same or different species or species or even an individual amino acid. It may also be a synthetic peptide. It may be a natural carbohydrate or other naturally occurring polymeric and biocompatible carbohydrates, such as polyols and amino acid sequences. Functional equivalence to natural apolipoprotein A-1, A-11 or A-iv may be conveniently determined using a lipid binding assay. The ability of the construct to exhibit substantially the same physiological response in a mammal may be conveniently determined by measuring the ability to perform the reverse transport of cholesterol in a test organism such as rabbits or rodents such as mice. The construct comprising the apolipoprotein and the heterologous radical is capable of performing reverse cholesterol transport as well as native apolipoproteins, or even better, despite the modification brought about by the addition of a heterologous radical. Preferably, the plasma half-life of the construct is higher compared to that of the wild-type apolipoprotein. The increase in the half-life may be due to the increased size of the apolipoprotein construct, which may reduce the rate of filtration through the kidneys, may be due to increased binding to HDL or may be due to a reduction in destruction of construction compared to native Apo A.

Preferably, the plasma half-life is at least double or triple, or at least four times, or at least 10 times. Likewise, binding affinity, such as the lipid binding affinity and (or the cholesterol binding affinity, of the construct is preferably increased compared to that of the wild-type apolipoprotein. Preferably, the binding affinity to lipids is increased by at least 5%, such as at least 10%, for example at least 15%, such as at least 20%, for example at least 25%, such as at least 30%, e.g. %, such as at least 50%, for example at least 75%, such as at least 100%, such as at least 150%, for example at least 200%, such as at least 300%. lipid binding affinity of the constructs according to the invention is equal to or less than the lipid binding affinity of the native apolipoprotein, the clinical effect may be increased due to the increase in the plasma half-life of the constructs according to the invention.

An increase in plasma half-life and / or an increase in lipid binding affinity have profound implications in the use of apolipoprotein constructs 7 for the treatment of atherosclerosis. The clinical effect of the apolipoprotein constructs of the present invention is thus expected to outweigh the effect of the wild type apolipoproteins. The invention also comprises analogs or variants of wild-type apolipoproteins that are capable of producing substantially the same physiological response in a mammal.

According to a second aspect, the invention provides an apolipoprotein construct having the general formula - apo AX, - wherein apo A is an apolipoprotein component selected from the group consisting of apolipoprotein AI, apolipoprotein All, apolipoprotein Aiv, an analog thereof or a variant thereof, and X is a heterologous radical selected from the group consisting of an oligomerization module and an apolipoprotein attached at the terminal.

According to another aspect, the invention provides a nucleotide sequence encoding an apolipoprotein construct, as defined above. Preferably, the nucleotide sequence is functionally linked to a regulatory sequence for the expression of the protein construct.

In another aspect, the invention provides a vector comprising the nucleotide sequence encoding the apolipoprotein construct and a transformed host cell comprising the nucleotide sequence as defined above. The apolipoprotein construct according to the present invention can be produced by different methods.

According to a first method, a transformed host cell is developed under conditions for promoting the expression of a protein construct according to the invention, which is encoded by a DNA insert in a construct, allowing to obtain and recover the protein construct and, optionally, processing the protein construct.

This method is the preferred method in the case where the entire construction is of a natural polypeptide and can thus be encoded by a corresponding nucleic acid sequence.

According to a second method, the construction of apolipoproteins can be produced by chemical synthesis of the heterologous radical and subsequent attachment of the radical to the apolipoprotein or to an analogue, obtaining an apolipoprotein construct, which is isolated and, optionally, subjected to new processing. This method is the preferred method in the case of the heterologous radical having a non-peptidic nature. However, there may also be conditions in which it is preferable to chemically synthesize the heterologous radical, such as in the case where it has a polypeptide nature. Such conditions may be that the heterologous radical is too short, such as less than 20 amino acids.

According to a third method, the construct of apolipoproteins can be produced by culturing a host cell modified under conditions to promote the expression of an apolipoprotein, or an apolipoprotein analog, encoded by a nucleic acid fragment and subsequent covalently attaching the apolipoprotein, or the apolipoprotein analog, to a heterologous radical to obtain an apolipoprotein construct, isolating the resulting apolipoprotein construct and, optionally, subjecting the construct to a further processing.

Finally, the construction of apolipoproteins can be produced by culturing a host cell transformed under conditions to promote expression of a protein encoded by a nucleic acid fragment encoding an oligomerization module and subsequently binding said module at least to an apolipoprotein to provide an apolipoprotein construct.

According to a further aspect, the invention provides a pharmaceutical composition comprising the construction of apolipoproteins, as defined above. Preferably, the pharmaceutical composition is capable of being administered parenterally, such as by injection. The invention also comprises the use of an apolipoprotein construct, as defined above, for the preparation of a pharmaceutical composition. The pharmaceutical composition may further comprise pharmaceutically acceptable excipients, adjuvants, additives, such as lipids, phospholipids, cholesterol or triglycerides. The pharmaceutical composition may be administered intravenously, intraarterially, intramuscularly, transdermally, pulmonary, subcutaneously, intradermally, intrathecally, through oral, anal, vaginal, conjunctival or intranasal tissue, by inoculation into tissue, such as tumor tissue, by means of an implant, or by oral route. The apolipoprotein construct as defined herein may also be used in gene therapy, wherein the DNA sequence encoding the apolipoprotein construct is used for transfection or infection in at least one cell population.

Detailed Description of the Invention The invention will now be described more fully with reference to the following figures. Figure 1 shows the amino acid sequence (in one-letter code) of human apolipoprotein A-I. Figure 2A shows the alignment of the multiple sequence CLUSTALW (1.74) of apolipoprotein A-1 using BLOSUM. The following sequences are aligned in the figure: apolipoprotein AI (Apo-AI) human precursor sp. P15568 | APA1_MACFA - Macaca fascicularis (macaque monkey) | APAl_HUMAN - Homo saplens (human) precursor apolipoprotein Al (Apo-AI) (Apo-AI) - Sus scrofa (bovine) apolipoprotein precursor AI (Apo-AI) of porcine sp P18648 | APA1_PIG (Apo-AI) of apolipoprotein AI (Apo-AI) from bovine sp | P15497 | APAl_BOVIN - (rabbit) apolipoprotein precursor apolipoprotein AI (Apo-AI) sp. P02648 | APAl_CANFA - Canis familiaris (dog) precursor apolipoprotein Al (Apo-AI) of rabbit sp | P09809 | APA1_RABIT - Oryctolagus cuniculus (rabbit) apolipoprotein precursor APA1_TUPGB - Tupaia glis belangeri (common tree muscaria) precursor of apolipoprotein AI (Apo-Al) spp Q00623 | APA1_M0USE - Mus musculus (mouse) precursor of apolipoprotein AI (Apo-AI) from rat sp | P04639 | APAl_RAT - Ratt US norvegicus (mouse) cholesterol transporter = AP0A-I, apolipoprotein AI (Apo-AI) from European hedgehog, Q9TS49 - Erinaceus europaeus (Western European hedgehog) apolipoprotein AI precursor (Apo-AI) AP02507 | APA1_C0TJA - Coturnix coturnix japonica (Japanese quail) apolipoprotein Ai (Apo-AI) precursor of domestic duck sp | P08508 | APA1_CHICK - Gallus gallus (chicken) apolipoprotein precursor AI (Apo-AI) APA1_ANAPL - Anas platyrhynchos (domestic duck) apolipoprotein precursor AI (APOA-I-1) of rainbow trout sp | 057523 | APll_ONCMY - Oncorhynchus mykiss (rainbow trout) (Salmo gairdneri) apolipoprotein AI precursor (Apo Salmon salmon (Atlantic salmon) apolipoprotein precursor AI (Apo-AI) of salmon Atlantic salmon (Apo-AI) of salmon Atlantic salmon (APA1_SALSA) - Salmo salar (Atlantic salmon) apolipoprotein precursor AI -AI) of zebrafish sp | 042363 | APA l_BRARE - Brachydanio rerio (zebrafish) (Zebra danio) apolipoprotein A-I precursor (Apo-AI) de oroada sp | 042175 | APAl_SPAAU - Sparus aurata (golden). Figure 2B shows aligned (single letter code) amino acid sequences of human apolipoprotein A-IV, monkey, mouse, baboon, pig and rat.

Figure 3: amino terminal amino acid sequence of the tetranectin region (SEQ ID NO 12). Amino acid sequence (in single-letter code) from EI to L51 of tetranectin. Exon 1 comprises residues E1 to D16 and exon 2 comprises residues VI7 to V49, respectively. The α-helix extends beyond L51 to K52, which is the C-terminal amino acid residue in the α-helix. Figure 4 shows an alignment of the amino acid sequences of the trimerizing structural element of the tetranectin family of proteins. The amino acid sequences (one-letter code) corresponding to residues V17 to K52 comprise exon 2 and the first three residues of exon 3 of human tetranectin; mouse tetranectin (Sorensen et al., Gene, 152: 243-245, 1995); tetranectin homologous protein isolated from reef shark cartilage (Neame and Boynton, 1992, 1996); and tetranectin homologous protein isolated from bovine cartilage (Neame and Boynton, accession number to the PATCHX database: u22298). Residues at positions a and d in heptad repeats are shown in bold. The listed consensus sequence of the trimerization structural element of the tetranectin family of proteins comprises residues at positions a and d of the heptase repeats shown in the figure and also other residues conserved in the region. The term " hy " designates an aliphatic hydrophobic residue. Figure 5 shows the plasmid pT7 H6UbiFx Apo A-I and their corresponding amino acid sequences. Expressed and processed polypeptide is comprised of amino acids 25 to 267 of human Apo AI (SEQ ID NO 1) and of the gly-gly residue attached thereto at the N-terminus. Figure 6 shows the plasmid pT7 H6UbiFx Cys-Apo AI and their corresponding amino acid sequences. Expressed and processed polypeptide consists of a N-terminal cysteine residue and amino acids 25 to 267 of human Apo AI (SEQ ID NO 2) and the gly-gly residue attached thereto at the N-terminus. Figure 7 shows the plasmid pT7H6 Trip-A-Apo AI-AmpR and their corresponding amino acid sequences. Expressed and processed polypeptide (SEQ ID NO 3) is composed of TTSE, a binding sequence, and amino acids 25 to 267 of human Apo A-I. Figure 8 shows the plasmid pT7H6 Trip-A-Apo A-i-del 433-AmpR and their corresponding amino acid sequences. Expressed and processed polypeptide (SEQ ID NO 4) is composed of TTSE, a binding sequence, and amino acids at 68 to 267 of human Apo A-1. Figure 9 shows the plasmid pT7H6FXCysApoAI and their corresponding amino acid sequences. Expressed and processed polypeptide consists of a N-terminal cysteine residue and amino acids 25 to 267 of human Apo A-I (SEQ ID NO: 2) and the gly-gly residue attached thereto at the N-terminus.

Figures 10A to G show illustrative examples of plasmids and corresponding amino acid sequences for apolipoprotein constructs according to the present invention.

Figure 10A: pT7H6-Trip-A-Apo AI K9A K15A: corresponds to pT7H6-Trip-A-Apo AI but two lysine residues in the trimerization region were mutated to remove heparin affinity. The mature protein product is designated Trip-A-AI K9A, K15A (SEQ ID NO 5).

Figure 10B: pT7H6 Trip-A-FN-Apo AI: corresponds to pT7H6-Trip-A-Apo AI, however, bases encoding the amino acid sequence SGH were inserted after the Trip A sequence and before the apo sequence THERE. The mature protein product is designated Trip-A-FN-AI (SEQ ID NO 6).

However, the BamHI site of pT7H6 Trip-A-FN-Apo AI was removed and the three sequences of the pT7H6 Trip-A-FN-Apo AI pT7H6 of inserted amino acids were altered such that the amino acid sequence between the trimerization sequence obtained from tetranectin and apo AI was changed from GSSGH to GTSGQ. The five amino acid sequence corresponds to a sequence in the fibronectin linker region. The mature protein product is designated Trip-A-FN-AI-final (SEQ ID NO 7).

Figure 10 D: pT7H6 Trip-A-FN-Apo AI-final K9AK15A: corresponds to pT7H6-Trip-A-FN-Apo AI-final, but the two lysine residues in the trimerization region were mutated to remove affinity with heparin. The mature protein product is designated Trip-A-FN-AI-final-K9A, K15A (SEQ ID NO 8).

Figure 10 E: pT7H6 Trip-A-TN-Apo AI: corresponds to pT7H6-Trip-A-Apo AI, however, bases encoding the KVHMK amino acid sequence were inserted after the Trip A sequence and before the sequence of apo AI. The mature protein product is designated Trip-A-TN-AI (SEQ ID NO 9). 15

Figure 10 F: pT7H6 Trip-A-TN-Apo AI-final: corresponds to pT7H6 Trip-A-TN-Apo AI, however, the BamHi site of TripT A-TN-Apo AI was removed in such a way that the amino acid sequence between the trimerization sequence obtained from tetranectin and apo AI was changed from GSKVHMK to GTKVHMK. The seven amino acid sequence corresponds to the tetranectin sequence followed by the trimerization domain. The mature protein product is designated Trip-A-TN-AI-final (SEQ ID NO 10).

Figure 10G: pT7H6 Trip-A-TN-Apo AI-final K9AK15A: corresponds to pT7H6-Trip-A-TN-Apo AI-final, but the two lysine residues in the trimerization region were mutated to remove heparin affinity. The mature protein product is designated Trip-A-TN-AI-final-K9A, K15A (SEQ ID NO 11).

Figure 10 H: pT7H6Fx-Hp (a) -ApoAI. The plasmid encodes the fusion protein between Hp (a) and ApoAI. The mature protein product is designated Hp (a) -ApoAI (SEQ ID NO 14). Figure 11 shows the result of binding of ApoA-1, TripA-ApoA-I and TripA-FN-ApoA-I to DMPC in the assay described in example 6. Figure 12 shows the binding of ApoA-I and TripA-ApoA the immobilized cubilin as described in example 7. Figure 13 shows the analytical gel filtration of Apo A-1, Trip-A-AI, Trip-A-TN-AI and Trip-A-FN-AI. As control BSA was included. The details are described in Example 5. Figure 14 shows the results of the evaluation of the disappearance of apolipoprotein A-I, TripA Apo-AI and the linker TripA-fibronectin to ApoA-I in plasma in mice. The experimental details are described in Example 8. 16

The functionality of the constructs according to the invention and the apo-A components of the constructs can be determined by means of a lipid binding assay, such as the DPCM assay described below. Furthermore, the in vivo effect on reverse cholesterol transport can be determined by administration to test animals, such as rabbits fed a high cholesterol diet, such as the method described by Miyazaki et al (Arteriosclerosis, Thrombosis , and Vascular Biology, 1995; 15: 1882-1888) or in Apo E-deficient mice (Sha PK et al, Circulation 2001, 103: 3047-3050).

Apolipoproteins or analogues

In the following text, the term " apo-A " means any apolipoprotein A, which comprises apolipoprotein A-1, apolipoprotein A-II or apolipoprotein A-IV, or any variant or analog thereof having the same lipid binding function.

Preferred apolipoprotein A-I analogues are those described in Figure 2A. Preferred apolipoprotein A-IV analogues are those described in Figure 2B.

As known variants of human Apo-AI sequences of figure 1 the following variants are referred to, the position of the change being indicated in relation to the sequence of figure 1, the change and, if appropriate, the name of the known variant. 27 P - > H (in MUNSTER-3C). 27 P - > R. 17 28 P - > R (in MUNSTER-3B). 34 R - > L (in BALTIMORE). 50 G - > R (in IOWA). 84 L - > R (in dominant amyloidosis 113 D -> E 119 A-> 1 D (in hita) 127 D -> N (in MUNSTER-3A) 131 missing (in MARBURG / MUNSTER-2) 131 K - > M 132 W -> R (in TSUSHIMA) 133 E -> K (in FUKUOKA) 151 R -> 3 (PARIS) 160 E -> K (in NORWAY) (In GIESSEN) 168 L -> R (in ZARAGOZA) 171 E -> V 189 P -> R 197 R -> C (in MILANO) 222 E - > K (in MUNSTER-4).

According to the invention, the term " apolipoprotein " is intended to include functional equivalents of at least one sequence of Figures 1, 2a and 2b, or a fragment of at least one sequence of Figures 1, 2a and 2b, which comprise a predetermined amino acid sequence. The term " fragment " means: i) fragments comprising an amino acid sequence that is capable of being recognized by an antibody also capable of recognizing the predetermined amino acid sequences in Figures 1, 2a or 2b and / or ii) fragments comprising an amino acid sequence that are capable of binding to a lipid, such as dimyristoyl phosphatidylcholine or cholesterol, and / or a receptor, which is also capable of binding to predetermined amino acid sequences in Figures 1, 2a or 2b.

According to the present invention, a functional equivalent of an apolipoprotein, or fragments thereof, may be obtained by addition, substitution or deletion of at least one amino acid. In the event that the amino acid sequence comprises a substitution of one amino acid for another, then such substitution may be a conservative amino acid substitution. Fragments of the sequence in Figures 1, 2a and 2b may comprise more than one such substitution, such as, eg, two conservative amino acid substitutions, for example seven or eight conservative amino acid substitutions, such as between 10 and 15 substitutions of conservative amino acids, for example, between 15 and 25 conservative amino acid substitutions, such as between 25 and 75 conservative amino acid substitutions, for example, between 75 and 125 conservative amino acid substitutions, such as between 125 and 175 conservative amino acid substitutions. Substitutions may be performed with any one or more predetermined amino acid groups.

Exemplary fragments include those comprising one or more conservative amino acid substitutions, including one or more conservative amino acid substitutions in the same predetermined amino acid group or several conservative amino acid substitutions, wherein each conservative amino acid substitution is generated by a substitution with a different group of predetermined amino acids.

Thus, a variant of the sequences of Figures 1, 2a or 2b, or fragments thereof according to the invention, may comprise, in the same sequence variant of Figures 1, 2a or 2b, or fragments thereof, or in different variants of the sequences of Figures 1, 2a or 2b, or fragments thereof, at least one substitution, such as several substitutions introduced independently of one another. The sequence variants of Figures 1, 2a or 2b, or fragments thereof, may then comprise conservative substitutions independently of one another, wherein at least one glycine (Gly) of said sequence variants of Figures 1, 2a or 2b, or fragments thereof of the sequences in Figures 1, 2a or 2b, is substituted with an amino acid selected from the group consisting of Ala, Val, Leu and Ile, and, independently, sequence variants of Figures 1, 2a or 2b, or fragments thereof, wherein at least one of said alanines (A1a) of said sequence variant of the Figures 1, 2a or 2b, or fragments thereof, is substituted with an amino acid selected from the group consisting of Gly, Val, Leu and Ile, and independently, a sequence variant of Figures 1, 2a or 2b, or fragments thereof, wherein at least one valine (Vai) of said sequence variant in Figures 1, 2a or 2b, or fragments thereof, is substituted Aa, and Ile, and independently sequence variants in Figures 1, 2a or 2b, or fragments thereof, wherein at least one of said leucine (Leu) is selected from the group consisting of Gly, Ala, Leu and Ile, of said variant of the sequences in Figures 1, 2a or 2b, or fragments thereof, is substituted with an amino acid selected from the group consisting of Gly, Ala, Val and Ile, and independently sequence variants in Figures 1, 2a or 2b, or fragments thereof, wherein at least one isoleucine (Ile) of said sequence variants in Figures 1, 2a or 2b, or fragments thereof, is substituted with an amino acid selected from the group consisting of Gly, Ala , Vai and Leu, and independently variants of the sequences in Figures 1, 2a or 2b, or fragments thereof, wherein at least one of said aspartic acids (Asp) of said sequence variants in Figures 1, 2a or 2b, or fragments thereof, is substituted with an amino acid selected from the group consisting of Glu, Asn and Gin, and, independently, sequence variants in Figures 1, 2a or 2b, or fragments thereof, wherein at least one of said phenylalanines (Phe) of said sequence variants in Figures 1, 2a or 2b, or fragments thereof, is substituted with an amino acid selected from the group consisting of Tyr, Trp, His, Pro, and preferably selected between the amino acid set Tyr and Trp, and, independently, sequence variants in Figures 1, 2a or 2b, or fragments thereof, wherein at least one of said tyrosines (Tyr) of said sequence variants in Figures 1, 2a or 2b or its sequence fragments in Figures 1, 2a or 2b is substituted with an amino acid selected from the group consisting of Phe, Trp, His, Pro, and preferably a (Arg) of said fragment of the sequences in Figures 1, 2a or 2b, or fragments thereof, wherein at least one of said arginines (Arg) of said fragment of the sequences in Figures 1a, 2a or 2b is substituted with an amino acid selected from the group consisting of Lys and His, and independently sequence variants in Figures 1, 2a or 2b, or fragments thereof, wherein at least one lysine (Lys ) of said sequence variants in Figures 1, 2a or 2b, or fragments thereof, is substituted with an amino acid selected from the group consisting of Arg and His, and, independently, sequence variants in Figures 1, 2a or 2b, or fragments thereof, wherein at least one of said asparagines (Asn) of said sequence variants in Figures 1, 2a or 2b, or fragments thereof, is substituted with an amino acid selected from the group consisting of amino acid sequence consisting of Asp, Glu and Gin, and independently sequence variants in Figures 1, 2a or 2b, or fragments thereof, wherein at least one glutamine (Gin) of said sequence variants in Figures 1, 2a or 2b, or fragments thereof, is substituted with an amino acid selected from the group consisting of Asp, Glu and Asn and, independently, sequence variants in Figures 1, 2a or 2b, or fragments thereof, wherein at least one proline (Pro) of said sequence variants in Figures 1, 2a or 2b, or fragments thereof, is substituted with an amino acid selected from the group consisting of Phe, Tyr, Trp and His, and, independently, sequence variants in the Figures 1a, 2a or 2b or fragments thereof, wherein at least one of said cysteines (Cys) of said sequence variants in Figures 1, 2a or 2b, or fragments thereof, is substituted with an amino acid selected from amino group consisting of Asp, Glu, Lys, Arg, His, Asn, Gin, Ser, Thr and Tyr.

Based on the foregoing, it is evident that the same variant, or a fragment thereof, may comprise more than one conservative amino acid substitution from more than one group of conservative amino acids, as defined hereinbefore. The addition or deletion of an amino acid may be an addition or suppression of 2 to 10 amino acids, such as 10 to 20 amino acids, for example, 20 to 30 amino acids, such as 40 to 50 amino acids. However, additions or deletions of more than 50 amino acids, such as additions of 10 to 200 amino acids, also fall within the scope of the present invention. More specifically, it is possible to remove 43 amino acids from the N-terminus from the sequence of Figure 1 without substantially altering the lipid binding effect of the protein. Such deletion is included in SEQ ID NO 4 with part of the apolipoprotein of the construct.

It will be appreciated that the invention relates to apolipoproteins comprising at least one fragment of the sequences of Figures 1, 2a or 2b which is capable of binding to lipids, such as DPMC, including any variants and equivalents of at least one single fragment .

In one embodiment, the apolipoprotein according to the invention, including any functional equivalents and fragments thereof, may comprise less than 243 amino acid residues, such as less than 240 amino acid residues, for example less than 225 residues of amino acid such as less than 200 amino acid residues, for example less than 180 amino acid residues, such as less than 160 amino acid residues, for example less than 150 amino acid residues, such as less than 140 amino acid residues, for example less than 130 amino acid residues, such as less than 120 amino acid residues, for example less than 110 amino acid residues, such as less than 100 amino acid residues, for example less than 90 amino acid residues, such as less than 85 residues of amino acid, for example less than 80 amino acid residues, such as less than 75 amino acid residues, for example less than 70 amino acid residues, t al as less than 65 amino acid residues, for example less than 60 amino acid residues, such as less than 55 amino acid residues, for example less than 50 amino acid residues.

Fragments

Particularly preferred is a fragment which comprises the lipid binding region of the native sequences of Figures 1, 2a or 2b. However, the invention is not limited to fragments comprising the lipid binding region. Also encompassed in the present invention are the deletions of such fragments which give rise to functionally equivalent fragments of the sequences in Figures 1, 2a or 2b which comprise less than the lipid binding region. Functionally equivalent peptides of the sequences of Figures Figure 1, 2a or 2b, and fragments thereof according to the present invention may comprise less or more amino acid residues than the lipid binding region. Preferably, the fragment comprises at least amino acids 100 to 186 of apo-A-I 24 or a variant thereof or a functional equivalent thereof. It has been determined that this central domain and the α-helices within the domain are directly involved in the interactions with the phospholipids. Thus, it is quite likely that this region plays an important role in the functional properties of apo-A-1.

In a preferred embodiment, " functional equivalence " as used in the present invention is set forth by reference to the corresponding functionality of a predetermined fragment of the sequences in Figures 1, 2a or 2b.

It will be apparent that the functional equivalents of the sequence variants of Figures 1, 2a or 2b will exhibit gradually different amino acid sequences of the preferred predetermined sequence as the number and extent of insertions, deletions and substitutions, including conservative substitutions, increases. This difference is measured as the reduction in homology between the preferred predetermined sequence and the fragment or functional equivalent.

All functional fragments or equivalents of apolipoprotein are within the scope of the present invention, regardless of the degree of homology they present to a predetermined preferred sequence of apolipoprotein. The reason for this is that some regions of the sequences in Figures 1, 2a or 2b are likely to be easy to mutate or to be completely suppressed without having any significant effect on the binding activity of the resulting fragment. 25

A functional variant obtained by substitution may exhibit some form or degree of native activity of the sequences in Figures 1, 2a or 2b, and yet be less homologous if amino acid side chain containing residues with identical functionality are substituted. In this regard, the term "identical functionality" refers to the dominant characteristics of the side chains, such as the hydrophobic, basic, neutral or acidic characteristics, or the presence or absence of steric material. Accordingly, in accordance with a variant of the invention, the degree of identity between i) a particular fragment of the sequences in Figures 1, 2a or 2b that is capable of producing an effect and ii) a preferred predetermined fragment is not the primary measure of the fragment as variant or functional equivalent of a preferred predetermined fragment of the sequences in Figures 1, 2a or 2b, according to the present invention. Homology between the amino acid sequences can be calculated by well known algorithms such as BLOSUM 30, BLOSUM 40, BLOSUM 45, BLOSUM 50, BLOSUM 55, BLOSUM 60, BLOSUM 62, BLOSUM 65, BLOSUM 70, BLOSUM 75, BLOSUM BLOSUM 85 or BLOSUM 90. Preferably, the algorithm used is BLOSUM 30.

Fragments that share at least some homology with the fragment of the sequences in Figures 1, 2a or 2b should be considered to fall within the scope of the present invention, if at least about 40% homologous with the apolipoprotein or a fragment thereof, such as at least about 50% homologous, for example at least about 60% homologous, such as at least about 70% homologous, for example at least about 75% homologous, such as at least about 80% homologous , for example at least about 85% homologous, such as at least about 90% homologous, for example at least 92% homologous, such as at least 94% homologous, for example at least 95% homologous, such as at least 96% % homologues, for example at least 97% homologues, such as at least 98% homologues, for example at least 99% homologous to the sequence fragment in Figures 1, 2a or 2b. According to a variant of the invention, percentages of homology designate percentages of identity.

Further factors which may be taken into account in determining functional equivalence according to the meaning used herein include i) the ability of antiserum against one of the sequences of Figures 1, 2a or 2b to detect fragments of the sequences in the figures 1, 2a or 2b according to the present invention, or ii) the suitability of a functionally equivalent fragment to compete with the sequences of Figures 1, 2a or 2b in a lipid binding assay.

Conservative substitutions may be introduced at any position of a preferred predetermined apolipoprotein or fragment thereof. However, it will also be desirable to introduce non-conservative substitutions, in particular, but not limited to, a non-conservative substitution at any one or several positions.

A non-conservative substitution which gives rise to the formation of a functionally equivalent fragment of the sequences in Figures 1, 2a or 2b would, for example, i) differ substantially in polarity, for example a residue with a non-polar side chain (Ala, Leu, Pro, Trp, Val, Ile, Leu, Phe or Met) substituted by a residue having a polar side chain, such as Gly, Ser, Thr, Cys, Tyr, Asn, or Gin, or by a charged amino acid, such as such as Asp, Glu, Arg or Lys, or by replacing a charged or polar residue with a nonpolar residue; and / or ii) differ substantially in their effect on the orientation of the principal structure of the polypeptide, such as the substitution of or with Pro or Gly by another residue; and / or iii) differ substantially in terms of electrical charge, for example by substitution of a negatively charged residue, such as Glu or Asp, by a positively charged residue, such as Lys, His or Arg (and vice versa); and / or iv) differ substantially in steric mass, for example by substituting a bulky residue, such as His, Trp, Phe or Tyr, by one with a minor side chain, eg Ala, Gly or Ser (and vice versa) versa).

In one embodiment, amino acid substitution may be effected based on their hydrophobicity or hydrophilicity values and on the relative similarity of amino acid side chain substituents, including charge, size, and the like. Examples of amino acid substitutions that take into account many of the above-mentioned characteristics are well known to those skilled in the art, including: arginine and lysine; glutamate and aspartate; serine and threonine; glutamine and asparagine and valine, leucine and isoleucine.

In addition to the variants described herein, it is possible to formulate sterically similar variants to copy the important portions of the variant structure, wherein such compounds may also be used in a manner identical to the variants of the invention. This can be achieved by means of modulation and chemical design techniques well known to those skilled in the art. It will be appreciated that such sterically similar constructs also fall within the scope of the present invention.

The component X

Preferably, the X component of the protein construct according to the invention is essentially non-immunogenic. For example, the component X may be an amino acid, a carbohydrate, a nucleic acid sequence, an inert protein or a polypeptide, which have virtually no effect and in particular no immunological effect on animals.

Preferably, the X component is non-immunogenic and does not negatively interfere with respect to ligand binding, i.e., the apolipoprotein component should not be directed to an undesired site through the interactions of the X component with the ligand.

In one embodiment, component X consists of only one amino acid, wherein said amino acid is preferably a cysteine residue, which may be placed at the N-terminus, at the C-terminus or internally at the apolipoprotein component. Such a construct may form a dimer with other identical or similar constructs. Preferably, a linker between the terminal cysteine residue and the apolipoprotein component is introduced to facilitate correct folding and interaction with lipids in the construct. 29

More preferably, however, component X comprises a peptide having more than 1 amino acid, such as more than 2 amino acids, for example more than 5 amino acids, such as greater than 10 amino acids, e.g. more than 15 amino acids such as greater than 20 amino acids, such as greater than 30 amino acids, for example greater than 40 amino acids, such as greater than 50 amino acids, for example greater than 75 amino acids, such as greater than 100 amino acids, for example more than 200 amino acids, such as greater than 300 amino acids, for example more than 400 amino acids, such as more than 500 amino acids, for example more than 600 amino acids, such as more than 700 amino acids, for example more than 400 amino acids, e.g. more than 800 amino acids, such as greater than 900 amino acids, for example greater than 1000, 1250, 1500, 2000 or 2500 amino acids.

In case the component X is a protein, then this protein is preferably a protein of a mammal and more preferably a human protein. Examples of suitable proteins are plasma proteins, such as serum albumin or albumin or other non-immunogenic peptide or protein, such as the plasminogen serine protease fragment or other serine proteases engineered to be inactivated by disruption of the catalytic triad; and the immunoglobulin heavy chain constant region. Most preferably, the protein comprises serum albumin. Even more preferably, the protein comprises an apolipoprotein which contains an amphipathic helix containing the apolipoprotein.

According to a particularly preferred embodiment of the invention, component X comprises an apolipoprotein component selected from the group consisting of apolipoprotein A-1, A-11, AIV, an analogue thereof, functional variant or fragment thereof. The two apolipoprotein components may be linked linearly or may be attached via a non-native terminal cysteine supplement bridge.

The higher oligomers and also the dimers of the apolipoprotein component which comprises at least one non-native cysteine residue can be prepared and linked through cysteine bridges under suitable conditions. Oligomers linked by disulfide bridges may be serially linked (apo-A-S-S-apo-A or apo-A-S-S-apo-A-S-S-apo-A or higher oligomers). The protein construct according to the invention may also comprise two, three or more apolipoproteins, or analogs thereof, serially or covalently linked to one another. This can be achieved by attaching the C-terminus of a first apolipoprotein to the N-terminus of the next apolipoprotein and so on. The proteins may also be ligated after transcription and translation or the nucleotide sequence may simply comprise two, three or more coding sequences for the construction of apolipoproteins in question, as well as optional linker peptides between the apolipoproteins.

Thus, it is possible to avoid the need for a heterologous radical to effect binding. It is expected that constructs having two, three or more apo-A units, practically all apo-A units will participate in lipid binding, thus contributing to the functionality of the building. Thus, these multi-apo-A constructs are expected to have increased lipid binding ability compared to native apo-A. It is an additional advantage of these constructs when compared to native apo A that they have an increased plasma half-life compared to native apo A.

Such constructs comprising more than one apolipoprotein component may have a combination selected from the following set: Dimers:

A-I A-I; A-II AII; A-IV A-IV; A-I-II; A-I-IV; A-II A-IV.

Trímeros: A-I A-II A-IV; A-I-A-II-A; A-I A-I A-I; A-I-A-IV; A-II A-II A-I; A-II A-II A-IV; A-II A-II A-II; A-IV A-IV A-IV; A-IV A-IV A-II; A-IV A-IV A-I. Oligomerization Modules

According to a particularly preferred variant of the invention, the heterologous radical is an oligomerization modulus. In this context, an oligomerization module is a peptide or a protein or a part of a protein that is capable of interacting with other, similar or identical oligomerization modules. The interaction is of the type that produces multimeric proteins or polypeptides. Such interaction may be caused by covalent bonds between the components of the multimer, as well as by hydrogen bonds, hydrophobic forces, van der Waals forces, salt bridges. The invention also encompasses oligomerization modules of a non-peptidic nature, such as an amino acid sequence of DNA, RNA, LNA or PNA. Techniques for the connection between proteins and nucleic acid sequences will be understood by a person skilled in the art. The oligomerization module may be a dimerization module, a trimerization module, a tetramerization module or a multimerization module.

In case the apolipoprotein or an analogous part of the construct is coupled to an oligomerization module, then it is possible to prepare multimers of the construct simply by mixing a solution of constructs (oligomerization module attached to the apolipoprotein part) under suitable conditions. By this means, it is possible to prepare dimers, trimers, tetramers, pentamers, hexamers or higher depending on the type of oligomerization module that is attached to the apolipoprotein part of the construct.

The multimers according to the invention may be homomers or heteromers, since different apolipoproteins may be attached to the oligomerization modules and incorporated into the multimer. It may be advantageous to mix different types of apolipoproteins in this way to obtain an improved clinical effect of the construct. Preferred homomers are Apo-A-I trimers and Apo-A-IV trimers.

According to a particularly preferred variant of the invention, the oligomerization module is obtained from tetranectin and, more specifically, comprises the trimerization structural element of tetranectin (hereinafter referred to as TTSE, SEQ ID NO 12), which is described further in WO 98/56906. The amino acid sequence of TTSE is shown in SEQ ID NO 12. The trimerization effect of TTSE is brought about by a spiral structure that interacts with the spiral structure of two other TTSEs to form a trimer, which is exceptionally stable. Another advantage of TTSE is that it is a weak antigen (WO 98/56906).

Preferably, the heparin binding site, which is located in the N-terminal region of exon 1 (Figure 4) is eliminated by removal or mutagenesis of the N-terminal lysine residues (residues 9 and 14 of SEQ ID NO 12) (Nielsen et al., 1997, FEBS Lett 412: 388-396), without inhibiting trimerization. Preferably, lysine residues are mutagenised to alanine. TTSEs which include most or all of exon 1 thus confer an affinity for sulfated polysaccharides to any protein designed to incorporate such TTSEs as part of their structure. However, if desired, it may be possible to reduce or eliminate this affinity by N-terminal truncation or mutagenesis of the lysine residues in the part of the TTSE corresponding to the N-terminal amino acid residues of tetranectin (Lorentsen et al 2000, Biochem J 347: 83-87 ). The interaction domain of the trimerization module according to the invention is preferably of the same type as the TTSE, namely a triple alpha helix coil. The TTSE may be from human tetranectin, rabbit tetranectin, mouse tetranectin or shark cartilage type C lectin. Preferably, the TTSE comprises a sequence having at least 68%, such as at least 75%, for example at least 81%, for example at least 87%, such as at least 92% identity with the consensus sequence of SEQ ID NO 12. Thus, TTSE analogs having substantially the same trimerization effect are also encompassed by the invention. 34

Preferably, the cysteine residue 50 of TTSE (SEQ ID NO 12) should be mutagenised to serine, threonine, methionine or any other amino acid residue to thereby prevent the formation of an undesired interchain disulfide bridge, which could lead to undesired multimerization. The presence of a trimer can be confirmed by well-known techniques, such as gel filtration, SDS-PAGE or native SDS gel electrophoresis, depending on the nature of the trimer. A preferred method for conferring the presence of an oligomer is to bind to DMSI (dimethylsubirimidate), followed by SDS-PAGE.

According to a preferred embodiment of the invention, the protein construct is obtained by the attachment of two or more apolipoproteins to oligomerization modules. The advantage of this variant is that the binding of individual apolipoproteins to each other has no place in the apolipoprotein but in the oligomerization module. Thus, the nature of the wild type apolipoprotein is conserved and the apolipoprotein retains the secondary and tertiary structure, which is an advantage for its physiological function. By further introducing a peptide spacer between the apolipoprotein and the oligomerization module it is ensured that both components of the construct can perform their interaction with lipids and with other oligomerization modules, respectively, without being affected by the interactions of the other component. Preferably, the peptide spacer is non-immunogenic and has an essentially linear three-dimensional structure. It is possible to carry out the oligomerization of different or equal apo-A units using an oligomerization module, such as a dimerization module, a trimerization module, a tetramerization module, a pentamerization module, a hexamerization module or a modulus multimerization. The oligomerization modules may comprise a spiral structure capable of recognition and interchain interaction. The general method for producing an artificial trimer of a protein or a peptide comprises identifying a trimerization module from proteins which form trimers in nature. Through careful analysis, the domain responsible for the protein-protein interaction can be identified, isolated and linked to the protein or peptide to be trimerized. According to the invention, such trimerization does not necessarily comprise the formation of an apolipoprotein trimer or an analogue. It is also possible to bind only one apolipoprotein to a trimerization module and to allow this peptide to trimerize with two other trimerization modules. Thus, the molecular weight of the apolipoprotein part and the half-life in plasma can be increased in comparison with the native apolipoprotein.

An example of an oligomerization module is disclosed in WO 95/31540 (HOPPE ET AL.) In which polypeptides comprising a collectin region are described. The amino acid sequence constituting the collectin region may be attached to any selected polypeptide. The trimers may be prepared under suitable conditions among three polypeptides comprising the amino acid sequence of the collectin region.

Another example of an oligomerization module is the haptoglobin al chain. The Î ± chain has a cysteine residue which can bind to another Î ± chain to form a dimer. A natural variant is the Î ± 2 chain, which has a part of the Î ± chain attached to the duplicated disulfide bridge. The Î ± 2 chain can form cysteine bridges with cysteine residues on other Î ± 2 or Î ± chains, thus forming trimers, tetramer, pentamer, hexamer and higher. In its natural form the α-chain is associated with a β-chain. It is possible to replace the β-chain with an apolipoprotein to produce an apo-A-chain (haptoglobin) construct.

Peptide Spacer The protein construct may also advantageously comprise a spacer moiety, which is covalently linked between the apolipoprotein, or an apolipoprotein analog, and the heterologous moiety. The effect of the spacer is to provide space between the heterologous radical and the apolipoprotein part of the construct. Thus, it is possible to guarantee that the secondary structure of the apolipoprotein part is not affected by the presence of the heterologous radical, thus maintaining the physiological effect of the apolipoprotein part. Preferably, the spacer is polypeptide in nature. Thus, the nucleic acid sequence encoding the spacer may be attached to the sequence encoding the apolipoprotein part of the construct and, optionally, the sequence to the heterologous radical, it being possible to produce the entire construct simultaneously. The design and preparation of suitable spacer moieties are known in the art and can conveniently be carried out by preparing fusion polypeptides having the apo-A-spacer-X format, wherein the spacer moiety is a polypeptide fragment ( often an inert fragment), to avoid undesired reactions between the spacer and neighboring parts or the construction. The spacer moiety may also be inserted between two TTSEs, allowing both to interact with a separate third TTSE to form a trimeric complex, which thus comprises two separate peptides: TTSE and TTSE-spacer-TTSE. This variant facilitates preparation of the apolipoprotein construct since most of the trimer, which only contacted with a dimer, can be synthesized with a single polypeptide comprising as fusion partners (apo-A designating any polypeptide sequence that forms the apolipoprotein part of the construct) apo-A-TTSE-spacer-TTSE-apo-A.

In variants in which two TTSEs are present in the same monomer it is preferred that the spacer moiety has a length and configuration which favors the formation of complexes involving the two TTSEs which are covalently linked by the spacer moiety. In this way, it is possible to reduce the problems resulting from the undesired formation of trimers with the formats (2 + 1 + 1), (2 + 2 + 2) and (2 + 2 + 1) (where only one of the TTSE of each monomer participates in complex formation).

Preferably, the peptide spacer comprises at least two amino acids, such as at least three amino acids, for example at least five amino acids, such as at least ten amino acids, for example at least 15 amino acids, such as at least 20 amino acids, for example at least 30 amino acids, such as at least 40 amino acids, for example at least 50 amino acids, such as at least 60 amino acids, for example at least 70 amino acids, such as at least 80 amino acids, such as at least 90 amino acids, such as 100 amino acids. The spacer may be attached to the apo-A and X components by means of covalent bonds, and preferably the spacer is essentially non-immunogenic, and / or is not prone to proteolytic cleavage and / or comprises no cysteine residue.

Likewise, the three-dimensional structure of the spacer is preferably linear or substantially linear. The following are examples of spacer sequences which are believed to be particularly preferred for attachment of apolipoprotein analogues to an X component. Preferred examples of peptide spacers or linkers are those which have been used to bind proteins without substantially impairing the function of the attached proteins or at least without substantially impairing the function of one of the bound proteins. More preferably, the linkers or spacers are used to bind proteins having spiral structures.

Linker based on tetranectin

The linkers may include tetranectin residues 53 to 56, which in tetranectin form a β chain, and residues 57 to 59 that form a curve in tetranectin 39 (Nielsen BB, Kastrup JS, Rasmussen H, Holtet TL, Graversen JH, Etzerodt M, Thorninger HC, Larsen IK, FEBS-Letter 412, 388-396, 1997). The sequence of the segment is GTKVHMK. This linker has the advantage that in native tetranectin bridging the trimerization domain with the CRD domain is assumed to be quite adequate to effect the binding of the trimerization domain to another general domain. Moreover, the resulting construct is not expected to be more immunogenic than building without a linker. The tetranectin based linker is rather preferable in case the component X comprises the TTSE.

Fibronectin-based linker The linker can be selected as a sub-sequence from the human fibronectin binding chain 3, which corresponds to amino acid residues 1992 to 2102 (SWISS-PROT numbering, entry P02751). Preferably, the subsequence is used: PGTSGQQPSVGQQ which encompasses amino acid residues 2037-2049, more preferred is the GTSGQ segment which corresponds to amino acid residues 2038 to 2042 of that subsequence. This construction is advantageous since it is known that it is not prone to proteolytic cleavage and is not expected to be quite immunogenic, since fibronectin is present at high plasma concentrations.

Linker derived from the superior hinge of human IgG3 The sequence of 10 amino acid residues obtained from the murine IgG3 superior hinge region, PKPSTPPGSS, was used for the production of antibodies dimerized by a spiral (Pack P. and Pluckthun, A. 40

Biochemistry 31, pp. 1579-1584 (1992)) and may be useful as a peptide spacer according to the present invention. Even more preferably, it may be the corresponding sequence obtained from the human upper IgG3 hinge region. Sequences derived from human IgG3 are not expected to be immunogenic to humans.

Flexible Linkers

Possible examples of flexible linker / spacer sequences are SGGTSGSTSGTGST, AGSSTGSSTGPGSTT or GGSGGAP. These sequences were used for the connection between designed spirals and other protein domains (Muller, K.M., Arndt, K.M. and Alber, T., Meth.Enzymology, 328, pp. 261-281 (2000).

The two components of the construction can be linked together by a covalent bond. This linkage may be formed between the X component and the C- or N-terminal amino acid of the apo-A component. The components may also be linked by means of more than one covalent bond. The covalent bond between the components may also comprise an S-S bridge, preferably between cysteine residues. These cysteine residues are placed at the C- or N-terminus of the apo-A component and at the terminus or internally at the X component.

Carbohydrates 41

In addition to the other components of the construction, the construction according to the present invention may comprise a carbohydrate moiety.

Structural trimerization element of tetranectin

A particularly preferred variant of the invention is the trimerization or partial trimerization of an apolipoprotein, or an analog thereof, with the trimerization module of tetranectin.

This technique is described in WO 98/56906 (TH0GERSEN ET AL.). The trimeric polypeptides are constructed in the form of monomeric polypeptide constructs comprising at least one tetranectin trimerization structural element (TTSE), which is covalently linked to at least one heterologous radical. The structural trimerization element of tetranectin is capable of forming a stable complex with two other trimerization structural elements of tetranectin. The expression " trimerization structural element " (TTSE) used in the present specification and claims is intended to denote the portion of a polypeptide molecule of the tetranectins family which is responsible for the trimerization between the monomers of the tetranectin polypeptides (SEQ ID NO 12). The term is also intended to encompass variants of TTSE from a naturally occurring member of the tetranectin family which have been modified in the amino acid sequence without adversely affecting, at any level, the trimerization properties relative to those presented by the tetranectin family member native. 42

Specific examples of such variants will be described more fully herein, it being generally preferred that the TTSE is obtained from human tetranectin, murine tetranectin, human or bovine cartilage type C lectin, or shark cartilage type C lectin. Particularly preferred are polypeptide monomer constructs having at least one TTSE obtained from human tetranectin. The sequence of 51 polypeptide residues encoded by tetranectin exon 1 and 2 (Figure 3, SEQ ID NO 12) appears to be unique for the tetranectin group of proteins (Figure 4), since it was not possible to establish homology sequence with other known polypeptide sequences. In the preparation for the experimental investigations of the tetranectin architecture a set of recombinant proteins was produced, wherein such set includes complete tetranectin, the CRD domain (which roughly corresponds to the polypeptide encoded by exon 3), a product corresponding to the polypeptide encoded by exons 2 + 3, a product corresponding to exons 1 + 2 (Holtet et al., 1996). Tetranectin is actually a trimer, but the polypeptide encoded by exon 2 is in fact capable of trimerizing by itself, as found from the observation that the recombinant protein corresponding to the 2 + 3 exons is in fact trimeric in solution. The analysis of the 3D structure of full length recombinant tetranectin crystals (Nielsen et al., 1996; Nielsen, 1996; Larsen et al., 1996; Kastrup, 1996) revealed that the polypeptide encoded by exon 2 plus three residues 43 encoded in exon 3 form a triple α-helix spiral structure. From the combination of sequence and structure data it becomes apparent that the trimerization in tetranectin is actually generated by a structural element (Figure 4), which comprises the amino acid residues encoded by exon two and by the first three residues of the exon 3 by means of an unusual heptate repeat sequence, which is apparently unique to tetranectin and other members of its group. This amino acid sequence (Figure 4) is characterized by two copies of the heptad repeats (abcdefg) with hydrophobic residues at positions a and d, as are other α-helix spirals. These two heptad repeats are followed in sequence by an unusual third copy of heptad repeat in which glutamine 44 and glutamine 47 do not replace only the hydrophobic residues at the aed positions but are directly involved in the formation of the spiral structure of triple α-helix. These heptad repeats are further flanked by two half-repeats with hydrophobic residues at positions d and a, respectively.

It is known that the presence of β-branched hydrophobic residues at positions a or d in the α-helix spiral influences the oligomerization state. Only one conserved valine (number 37) is present in the structural element of tetranectin. At position 29 of the tetranectin sequence no particular aliphatic residue seems preferable.

Briefly, the triple-stranded spiral structure in tetranectin is believed to be governed to a large extent by interactions which are unexpected in view of the characteristic interactions of the group of known spiral proteins.

TTSEs form surprisingly stable trimeric molecules. Experimental observations that (1) a substantial part of the recombinant proteins exist in the oligomeric state and can be crosslinked with trimer molecules even at 70øC and that (2) the exchange of monomers between different trimers can only be detected upon exposure to temperatures are evidence of a trimerization structural element of tetranectin with extremely high stability. This characteristic will be reflected in the amino acid sequence of the structural element. In particular, the presence and position of the glutamine-containing repeat in the sequential alignment of the heptad repeat, together with the presence and relative position of other conserved residues in the consensus sequence (Figure 4), is considered important for the formation of these stable trimeric molecules. For the most practical uses, the cysteine residue 50 should be mutagenized to serine, threonine, methionine or any other amino acid residue, to avoid the formation of an undesired interchain disulfide bridge, which may result in uncontrolled multimerization, aggregation, and precipitation of a polypeptide product that supports this sequence.

In particular, in conjunction with polypeptide encoded by trimer stabilization exon 1, the trimerization structural element of tetranectin is a truly autonomous polypeptide module which retains its structural integrity and propensity to generate a fairly stable homotrimeric complex, whether bonded or not via a peptide bond at one or both ends to other proteins.

This unique property is demonstrated by the fact that the polypeptide sequences obtained from heterologous proteins can be easily trimerized if they are attached, as fusion proteins, to the structural trimerization element of tetranectin. This remains valid regardless of whether the heterologous polypeptide sequences are placed at the amino or carboxy terminus of the trimerization element which allows the formation of a molecular assembly containing up to six copies of a particular polypeptide sequence or functional entities, or the formation of a molecular assembly containing up to six different polypeptide sequences, each contributing to its individual functional property.

Since three naturally occurring tetranectin TTSEs form a triple α-helix spiral, it is therefore preferred that the stable complex formed by the TTSEs of the invention also forms a triple α-helix spiral. The " tetranectin family " are polypeptides that share the consensus sequence shown in Figure 4 or a sequence that is homologous to the sequence level with this consensus sequence.

Thus, monomeric polypeptide constructs of the invention comprising a polypeptide sequence having at least 68% sequence identity with the consensus sequence shown in Figure 4 are preferred, although higher sequence identities are preferred, such as at least 75%, at least 81%, at least 87% and at least 92%.

Trip-A module

In the expression of plasmids according to the present invention, the TTSE (SEQ ID NO 12) modulus was modified as indicated by substituting Cys 50 for Ser and for inclusion of a lysine residue at the C-terminus. An SPGT sequence was added to the terminus N. This is a connection sequence to the trimerization module. The sequence has been inserted since it provides the opportunity to cut the DNA strand with Bgl II and Kpn K. A C-terminal linkage is provided at the C-terminus, which provides an opportunity to cut with Bam HI. This modified TTSE is designated by TripA and is described in SEQ ID NO 13. The trimerization module of the Apo A construct may thus advantageously comprise this sequence or a sequence which possesses at least 68% sequence identity with the sequence SEQ ID NO 13, with higher sequence identities being preferred, such as at least 75%, at least 81%, at least 87% and at least 92%.

Specific examples of constructs comprising the Trip A module are described in the examples.

Examples of constructs according to the invention The invention comprises the specific sequences described in the attached examples, such as SEQ ID NOs 2 to 11 and SEQ ID NO 14. Preferably the invention comprises SEQ ID NOs 3 to 11 and SEQ ID NO 14. Sequences that share at least 60% sequence identity, such as at least 70% identity sequence with these sequences are also within the scope of the invention, preferably sequences which share at least 80% of sequence identity, more preferably at least 90%, more preferably at least 95% and still more preferably at least 98%.

Production of protein construct

In order to produce a peptide component of the protein construct the cDNA encoding this part is inserted into an expression vector and transformed into a host cell. The host cell referred to above (which also forms part of the invention) may be prepared by conventional genetic manipulation techniques, which comprise insertion of a nucleic acid fragment (usually a DNA fragment) encoding the part of the polypeptide of a constructing a polypeptide monomer of the invention in a suitable expression vector, transforming a suitable host cell with the vector and culturing the host cell under conditions that allow the expression of the polypeptide portion of the polypeptide monomer construct. The nucleic acid fragment encoding the polypeptide may be placed under the control of a suitable promoter, which may be an inducible or constitutive promoter.

Depending on the expression system, the polypeptide can be recovered from the extracellular phase, the periplasm or the cytoplasm of the host cell.

Suitable vector and host cell systems are well known in the art, as can be seen from the vast amount of literature and materials available to one skilled in the art. Since the present invention also relates to the use of nucleic acid fragments of the invention in the construction of vectors and host cells, the following text provides a general description of such use and particular considerations about the practice of this aspect of the invention.

In general, prokaryotes are preferred for the initial cloning of nucleic sequences of the invention and for constructing vectors useful for the invention. For example, in addition to the particular strains mentioned hereinafter more fully below, there may be mentioned, by way of example, strains such as strain 294 of E. coli K12 (ATCC No. 31446), E. coli B and E. coli X 1776 (ATCC No. 31537). It is clear that these examples are merely illustrative and not limiting. Prokaryotes are also preferable for expression, since efficient purification techniques and protein refolding strategies are available. The above-mentioned strains, as well as E. coli W3110 (F-λ, prototrophic, ATCC No. 273325), bacillus, such as Bacillus subtilis, or other enterobacteria, such as Salmonella typhimurium or Serratia marcesans, and several species of pseudomonas.

Generally, plasmid vectors containing replication and control sequences, which are obtained from species compatible with the host cell, are used in conjunction with these hosts. The vector normally supports a replication site, as well as marker sequences that are capable of providing a phenotypic selection in the transformed cells. For example, E. coli is typically transformed using pBR322, which is a plasmid obtained from an E. coli species (see, e.g., Bolivar et al., 1977). Plasmid pBR322 contains genes for ampicillin and tetracycline resistance, thus providing a simple means for the identification of transformed cells. The plasmid pBR, or other microbial plasmid or phage, should also contain, or be modified to contain, promoters which may be used by the microorganism for expression.

The promoters commonly used in the construction of recombinant DNA include B-lactamase (penicillinase) and lactose promoter systems (Chang et al., 1978; Itakura et al., 1977; Goeddel et al., 1979) and a promoter system of tryptophan (trp) (Goeddel et al., 1979; EPO Appl. Publ. No. 0036776). Although these are the commonly used promoters, other promoters have already been discovered and used, details having been published concerning their nucleotide sequence, which allows one skilled in the art to functionally link them with plasmid vectors (Siebwenlist et al. 1980). There are certain prokaryotic genes which can be efficiently expressed in E. coli from their own promoter sequence, thus avoiding the addition of another promoter by artificial means.

In addition to prokaryotes, it is also possible to use eukaryotic microbes, such as yeast cultures. Saccharomyces cerevisiae, or common baker's yeast is the commonly used eukaryotic micro-organism, although there are several other commonly available strains. For example, for expression in Saccharomyces, the plasmid YRp7 (Stinchcomb et al., 1979; Kingsman et al., 1979; Tschemper et al., 1980) is commonly used.

This plasmid already contains the trp1 gene, which provides a selection marker for a mutant strain of yeast lacking the ability to grow in tryptophan, for example, ATCC No. 44076 or PEP4-1 (Jones, 1977). The presence of the trp alteration as a feature of the genome of the yeast host cell then provides an environment for detecting transformation through growth in the absence of tryptophan.

Suitable promoter sequences in yeast vectors include promoters for 3-phosphoglycerate kinase (Hitzman et al., 1980) or other glycolytic enzymes (Hess et al., 1968; Holland et al., 1978), such as enolase , glyceraldehyde-3-phosphate dehydrogenase, hexokinase, pyruvate decarboxylase, phosphofructo kinase, glucose-6-phosphate isomerase, 3-phosphoglycerate mutase, pyruvate kinase, triosephosphate isomerase, phosphoglucose isomerase and glycine kinase. In constructing suitable expression plasmids, the termination sequences associated with these genes will also be linked to the 3 'expression vector of the desired sequence to be expressed to obtain polyadenylation of the mRNA and termination. As other promoters having the additional advantage of having a transcription controlled by developmental conditions, the promoter region for alcohol dehydrogenase 2, isocytochrome C, acid phosphatase, degradation enzymes associated with nitrogen metabolism, the above-mentioned glyceraldehyde- 3-phosphate dehydrogenases and the enzymes responsible for the use of maltose and galactose. Any plasmid vector containing a yeast-compatible promoter sequence, origin sequence and end-termination sequence is suitable.

In addition to the microorganisms, it is also possible to use cell cultures obtained from multicellular organisms as hosts. In principle, any culture of cells, whether vertebrate or invertebrate, can be used. However, vertebrate cells are more interesting in that the propagation of vertebrates in culture (tissue culture) has become a routine procedure (Tissue Culture, 1973). Examples of useful host cell lines include VERO and HeLa cells, Chinese hamster ovary (CHO) cell lines and W138, BHK, COS-7 293 and MDCK cell lines.

Expression vectors for such cells usually include (if necessary) an origin of replication, a promoter located ahead of the gene to be expressed, in addition to any necessary ribosomal binding sites, RNA cleavage sites, a polyadenylation site and transcriptional termination sequences.

For use in mammalian cells, the control functions in expression vectors are often provided by viral materials. For example, promoters obtained from polyoma, adenovirus 2 and more frequently from Simian Virus 40 (SV40) are commonly used. Early and late SV40 virus expression promoters are particularly useful since they are easily obtained from the virus as a fragment which also contains SV40 viral origin of replication (Fiers et al., 1978). It is also possible to use shorter or longer SV40 fragments, as long as they include the approximately 250 bp sequence that extends from the HindIII site to the Bgl site, located at the viral origin of replication. In addition, it is also possible, and often desirable, to use promoter or control sequences normally associated with the desired gene sequence, as long as such control sequences are compatible with host cell systems.

An origin of replication may be provided by the construction of the vector to include an exogenous origin, such as can be obtained from SV40 or another virus (eg, Polyoma, Adeno, VSV, bpv) or may be provided by mechanism of host cell chromosome replication. In case the vector is integrated into a chromosome of the host cell, then the latter is often sufficient.

During the production of the polypeptide monomer constructs it may be necessary to further process the polypeptides, eg by introducing non-protein functions into the polypeptide, subjecting the material to suitable folding conditions (eg, using the strategies commonly used as suggested in WO 94/18227) , or by cleaving unwanted peptide radicals from the monomer (eg, expression enhancing peptide fragments which are undesired in the final product). In light of the above, the methods for the recombinant production of the polypeptide monomer construct of the invention are also encompassed by the invention, such as vectors which support and / or are capable of replicating the nucleic acids according to the invention in a cell host or in a cell line. According to the invention, the expression vector may be, e.g., a plasmid, a cosmid, a minichromosome or a phage. Of particular interest are vectors which are integrated into the host cell / cell line genome after introduction into the host.

Another part of the invention is transformed cells (useful in the methods described above) that support and are capable of replicating nucleic acid fragments of the invention; the host cell may be a microorganism, such as a bacterium, a yeast or a protozoan, or a cell obtained from a multicellular organism, such as a fungus, an insect cell, a plant cell or a mammalian cell. Of particular interest are the cells of the bacterial species Escherichia, Bacillus and Salmonella, the bacterium being preferred to E. coli.

Another part of the invention relates to a stable cell line producing the polypeptide portion of a construct according to the invention, and preferably such a cell line supports and expresses a nucleic acid of the invention.

Receptor binding The performance of the constructs according to the invention can be analyzed by determining the ability of the constructs to bind to the receptors or HDL proteins which can bind to the native apolipoproteins Al, A-II or A-54 IV . Such receptors and proteins include, but are not limited to, cubilin, megalin, class 1 depuration receptor B (SR-B1), ATP binding cassette 1 (ABC1), lecithin: cholesterol acyltransferase (LCAT) , cholesteryl ester transfer protein (CETP), phospholipid transfer protein (pltp). The dissociation constant, Kd, of the complex between cubilin and native apolipoprotein A I is 20 nM. It has been experimentally determined that an apolipoprotein A trimer I according to the present invention binds even more strongly to cubilin (Figure 12).

Affinity Markers The protein construct according to the invention may also comprise an affinity marker useful for purification of the construct. Preferably, such a label comprises a polyhistidine sequence. This sequence may advantageously be used for purification of the product on a Ni 2+ column, which will bind the polyhistidine sequence and hence the entire protein. Upon elution from the column, the polyhistidine sequence can be removed by cleavage by means of a proteinase, such as thrombin, which recognizes a specific sequence introduced in the construct between the protein construct and the polyhistidine sequence.

Other examples of affinity labels include, but are not limited to, well-known labels, such as an antigenic marker or a GST marker. It is possible to insert a proteolytic cleavage site between the label and the construct to cleavage the label. 55 Signal peptides

When expressing the constructs according to the invention in E. coli or a yeast, it may be preferred to include a signal peptide in the expression construct to ensure that the expressed protein is separated and can be harvested from the medium surrounding the cells, rather than using a more complicated process for isolating the expressed protein from within the cells. Specific examples of signal peptides for expression in yeast and E. coli which may be used in conjunction with the present invention include those described in WO 90/12879 (Sirtori et al), which describes a peptide of signaling for expression of Apo-Al and Apo-AIM in a yeast, and in WO 94/13819 (Kabi Pharmacia), which describes a signal peptide for the expression of Apo-AI and Apo-AIM in E. coli.

Production of apo-A-TTSE

To produce a construct comprising an apolipoprotein part and a TTSE, the cDNA encoding the apolipoprotein part is attached at the 3 'end to the 5' end of the cDNA encoding the TTSE. It is also possible to connect other units of TTSE and apolipoprotein units. A sequence encoding an enzyme cleavage site is further linked to the 3 'end of the sequence encoding the TTSE and, finally, a sequence encoding the polyhistidine is also attached. This can be done by conventional PCR techniques. The combined cDNA is inserted into an expression vector and transformed into the host cell.

Following expression in E. coli, the polyhistidine sequence is used to capture the heterologous protein on a Ni2 + column. After elution, the polyhistidine tail can be removed by a proteinase, such as Fx which cleaves the heterologous protein at the specific site that has been inserted between the TTSE and the polyhistidine sequence. The resulting apo-A-TTSE peptide can then be further processed by trimerizing itself to identical or different other apo-A-TTSE peptides. To improve expression in E. coli it may be advantageous to express the construct in the form of a fusion protein together with, e.g., ubiquitin, which may then be removed by cleavage.

Use of an apo-A construct for the preparation of a pharmaceutical composition The apo-A construct can be used for the preparation of a pharmaceutical composition. The composition may comprise pharmaceutically acceptable excipients, adjuvants, additives, such as phospholipids, cholesterol or triglycerides. The pharmaceutical composition may be administered intravenously, intraarterially, intramuscularly, transdermally, pulmonary, subcutaneously, intradermally, intrathecally, through oral, anal, vaginal, conjunctival or intranasal tissue by inoculation into the tissue, such as tumor tissue, for implant or oral.

Preferably, the formulation of the pharmaceutical compositions according to the invention is effected by means of techniques well known to those skilled in the art. This formulation may comprise pharmaceutically acceptable excipients, adjuvants, additives, such as phospholipids, cholesterol or triglycerides.

Administration of apo-A construction

The apo-A constructs according to the invention may be administered for the prevention and / or treatment of diseases associated with cholesterol, phospholipids and triglycerides, LDL and HDL disorders such as hypercholesterolemia, and arteriosclerotic diseases , such as atherosclerosis and myocardial infarction. Other indications are angina pectoris, angina pectoris, unstable angina pectoris, arterial stenoses, such as carotid stenosis, claudication, or cerebral arterial stenosis. In addition, the apolipoprotein constructs can also be used for the removal of endotoxins.

In one embodiment, administration comprises administering at least 50 mg of the construct each week to achieve a plasma concentration of approximately 0.5 g / L. Preferably, the construct is administered parenterally, such as by injections, suppositories, implants, etc. Preferably, the composition is administered in an amount comprising at least 50 mg of apolipoprotein construct per week, such as at least 100 mg / week, for example at least 250 mg / week, such as at least 500 mg / week, for example at least 750 mg / week, such as at least 1000 mg / week, for example at least 1250 mg / week, such as with at least 1500 mg / week, for example at least 2000 mg / week, such as at least 2500 mg / week, for example at least 5000 mg / week. The administration can be done daily, every two or three days, weekly, every two weeks, every three weeks or every four weeks.

In another embodiment, the construct is administered one, two or three times in much higher amounts particularly for the acute treatment of angina pectoris and angina pectoris or unstable angina pectoris. Administration may be effected for 1, 2, 3, 4, 5, 6, 7, 8 or up to 10 days. The amounts may be at least 10 mg / kg body weight, such as at least 20 mg / kg body weight, for example at least 30 mg / kg, such as at least 40 mg / kg, for example at least 50 mg / kg, such as at least 60 mg / kg, for example at least 70 mg / kg, such as at least 75 mg / kg, for example at least 90 mg / kg, such as at least 100 mg / kg, for example at least 125 mg / kg, such as at least 150 mg / kg, for example at least 200 mg / kg, such as at least 250 mg / kg, for example at least 300 mg / kg, such as at least 400 mg / kg, for example at least 500 mg / kg, such as at least 600 mg / kg, e.g. as at least 800 mg / kg, for example at least 900 mg / kg, such as at least 1000 mg / kg.

The constructs can also be administered orally. For this route of administration, the technology described in WO 99/46283, US 5,922,680, US 5,780,434 or US 5,591,433, US 5,609,871 or US 5,783,193 may be applied to the protein constructs according to the present invention. 59

Cell Population The invention also comprises the use of the nucleotide sequence according to the invention for gene therapy.

The genes may be transferred to a population of macrophages and subsequently transferred to a patient in need of such treatment. Thus, the transient expression is obtained, since macrophages have a limited life span in the blood vessels. Permanent transfection can be achieved by transforming liver cells. In the following, the invention is described by way of specific examples of variants of the invention, which are to be considered illustrative and non-limiting examples. The design of other constructions according to the invention is well known to those skilled in the art.

Example 1: Cloning of Άρο A-I

The cDNA encoding Apo A-1 was amplified from a human liver cDNA library (Clontech), using standard PCR techniques. For the Ubi-AI construct the primers were used: 5'-CAC GGA TCC ATC GAG GGT AGG GGT GGA GAT GAA CCC CCC CAG AGC-3 'and 5'-TCC AAG CTT ATT ACT GGG TGT TGA GCT TCT TAG TG- 3 '. The product was cloned into vector pT7H6Ubi (described by Ellgaard L. et al., Eur. J. Biochem., 1997; 244 (2): 544-51) using the Bam HI and Hind III cloning sites. For the construction of Trip-A-I the primers were used: 5'-AAG GGA TCC GAT GAA CCC CCC CAG AGC CCC-3 'and 5'-TCC AAG CTT ATT ACT GGG TGT TGA GCT TCT TAG TG-3'. The PCR product was cloned into the vector pT7H6tripa, described in WO 98/56906, using the Bam HI and Hind III cloning sites. For the construction of Trip-Al-del43 the primers were used: 5'-AGG GGA TCC CTA AAG CTC CTT GAC AAC TGG G-3 'and 5'-TCC AAG CTT ATT ACT GGG TGT TGA GCT TCT TAG TG -3' . The PCR product was cloned into the vector pT7H6tripa, described in WO 98/56906, using the Bam HI and Hind III cloning sites. For the construction of Ubi-Cys-AI primers were used: 5'-GGT GGA TCC ATC GAG GGT AGG GGT GGA TGT GAT GAA CCC CCC C-3 'and 5'-TCC AAG CTT ATT ACT GGG TGT TGA GCT TCT TAG TG-3 '. The product was cloned into vector pT7H6Ubi, described by Ellgaard L. et al. Biochem. 1997; 244 (2): 544-51), using the Bam HI and Hind III cloning sites. The plasmids generated are shown in Figures 4, 5, 6 and 7.

Expression of apolipoprotein A-1 (apo A-I) in E. coli Ubi-A-I and Trip-A-I, as well as other constructs described in the figures are conveniently expressed in E. coli AV-1 cells (Stratagene Inc.). Other cell lines can also be used. Cell culture and the induction of expression were performed as described for tetranectin in WO 98/56906.

Example 3: Isolation and Processing of Protein

The crude protein was isolated by phenol extraction as described for tetranectin in WO 98/56906. The redissolved granules were centrifuged from 6 liters of expression culture to remove the undissolved material, and the batch adsorption was then carried out in 50 ml of Ni2 + -NTA-Sepharose, which was prepared as described in Example 1.

as described in WO 98/56906. The column material was loaded onto a column and then washed with 500 mL of 8 M urea, 500 mM NaCl, 50 mM Tris-HCl, pH 8.0, and then with 200 mL of 6 M guanidinium HCl, Tris 50 mM HCl, pH 8.0, and finally with 300 mL of 500 mM NaCl, 50 mM Tris-HCl, pH

8.0. The protein was eluted with 500 mM NaCl, 50 mM Tris-HCl, pH 8.0, and 10 mM EDTA. 0.5 mg of Factor Xa was added to the protein and digested overnight at room temperature. It is also possible to use thrombin for this purpose. The protein was subjected to gel filtration on a G-25 sephadex column (Pharmacia) in 500 mM NaCl buffer, 50 mM Tris-HCl pH 8.0. The undigested protein was removed by passage of the protein solution on a Ni2 + -NTA-Sepharose column, prewashed with 500 mM NaCl, 50 mM Tris-HCl pH 8.0, and then washed with 500 mM NaCl , 50 mM Tris-HCl, pH 8.0. Undiluted protein eluted with 500 mM NaCl, 50 mM Tris-HCl, pH 8.0, and 10 mM EDTA. Further purification can be performed using a Sp-Sepharose ion exchange column.

Example 4: Removal of lipids from proteins

The proteins were gel-filtered in a 10 mM (nH 4) 2 CO 3 solution, pH 8.8, and lyophilized. Lyophilized proteins were resuspended in 25 ml of a 1: 1 mixture of methanol / chloroform cooled, incubated on ice for 30 minutes and centrifuged at 3000 g for 20 minutes. The pellets were resuspended in 25 mL of a 1: 2 mixture of methanol / chloroform cooled, equilibrated for 30 minutes on ice and centrifuged again. The supernatant was removed, the pellets were rapidly air-dried and then redissolved in 6 M guanidinium HCl, 50 mM Tris-HCl pH 8.0 overnight.

Example 5: Multimerization test

Crosslinking Multimerization can be determined by cross-linking the multimer and subsequent SDS-PAGE analysis.

60 μl of a 0.2 mg / ml protein was dissolved in 150 mM Na-borate, pH 9.0 equilibrated to the desired temperature for 30 minutes, 5 μl of 20 mg / ml dimethylsuberimidate was added and the incubate for 30 minutes at the desired temperature. The crosslinking was quenched by the addition of 5 μl of 3 M Tris-HCl, pH 9.0. Dimethylsuberimidate causes the lysine residues located within a short distance from each other to form a covalent bond. As a result it is obtained that the proteins that had formed multimer are linked together. The molecular weight of the multimer can be estimated by subsequent SDS-PAGE.

The cross-linking products were analyzed by SDS-PAGE on 8% -16% polyacrylamide gels. Optionally, an adjuvant, such as a lipid, was incubated in the cross-linking mixture, in which case the protein was preincubated with the adjuvant.

Filtration through analytical gel Multimerization can also be determined by analytical gel filtration. 63

The protein was dissolved in 500 mM NaCl buffer, 50 mM Tris-HCl pH 8.0, and subjected to gel filtration on a Superdex 200 HR 10/30 column with the desired buffer at room temperature and with a flow rate of 0.25 ml / minute. For standard procedures the buffer was 100 mM NaCl, 50 mM Tris-HCl pH 8.0. From Figure 13, it is possible to observe that Apo A-I elutes when the composite has a peak, the main ones being centered approximately 14.5 mL and 16.5 mL. BSA, with a molecular weight of 68 kDa, elutes to approximately 14.5 mL, which indicates that the Apo AI peak at 16.5 mL corresponds to monomeric Apo AI, while the other major peak corresponds to complexes of own association of apo AI. The constructs fused to the trimerization domain will all elute with a major peak at approximately 10.7 mL and a secondary peak at 14 mL. The 14 mL peak probably corresponds to the trimeric form of the constructs, while the main peak at 10.7 mL corresponds to a product with a higher molecular weight. The product is presumed to be formed by association of the trimers. This indicates that the fusion of apo AI with the trimerization domain does not give rise only to trimers, but also to the formation of large complexes, in which Apo Ai units may interact with other AI units, i.e. native apo AI by interacting with other Apo AI molecules.

Example 6: kinetics of the association of the protein construct with dimyristoyl phosphatidylcholine (DMPC) The ability of the constructs according to the invention to bind to lipids can be conveniently determined using a well known assay such as the combination with dimyristoyl phosphatidylcholine (DPMC). The assay was performed as described by Bergeron J. et al. (1997), Biochem. Biophys. Acta, 1344, 139-152. Anhydrous DPMC suspension in 100 mM NaCl, 50 mM Tris-HCl, pH 8.0, and 0.25 mM EDTA were placed in suspension at a temperature above the transition temperature and at a concentration of 0.5 mg / mL. The protein sample, the buffer and the DMPC suspension were incubated at 24øC for 10 minutes and then mixed such that the final concentration of DMPC was 0.4 mg / ml, with a protein concentration equal to 5.2 μΜ (of monomer). After the turbidity reduction of the blend, reflecting an increase in the lipid-protein association, the absorbance of the blend was determined at 325 nm. The assay was performed four times for each apo AI, Trip-A-AI and Trip-A-FN-AI and once in the absence of protein addition. From Figure 11, it can be seen that all of the Apo A-1 constructs bind to DMPC. For all three tested constructs the turbidity disappeared completely after 24 hours, indicating that the ability of the fusion proteins to bind to DMPC is present in the fusion proteins. However, both Trip-A-Apo AI and Trip-A-FN-Apo AI bind more slowly to DMPC than native Apo AI at 24 ° C, which is the only temperature at which the assay is functional .

Example 7: Surface plasmon resonance analysis of the binding of the derivatives to cubilin

The assay was performed as described by Kozyraki R, Fyfe J, Kristiansen M, Gerdes C, Jacobsen C, Cui S et al. The intrinsic factor-vitamin B12 receptor, cubilin, is a high-affinity apolipoprotein a-i receptor facilitating endocytosis of high-density lipoprotein. Nat Med 1999; 5 (6): 656-661. The concentration of the apolipoprotein construct used was 0.5 μΜ. Only the results (Figure 12) for TripA-AI and for apo A-1 are presented. For TripA-AI a binding similar to that observed for TripA-FN-AI and TripA-TN-AI was observed. There was increased response with the trimerization of apo A-I, in particular there was a decrease in the dissociation, based on " avidity " gains from the interaction of a multimer with an immobilized target as compared to the monomer (multivalence bonus). This shows that apo A-I was able to bind to cubilin in the trimeric state and that more than one apo A-I had a configuration capable of interacting with the cubilin, which indicates a correct folding of the apo A-I unit in the trimeric construction.

Example 8: evaluation of the disappearance of apolipoprotein AI, TripA Apo-AI and TripA fibronectin-linker Apo-AI in mice In mice in three groups, each with five mice, was injected 1 mg of apo AI, Trip-A -AI or Trip-A-FN-AI, respectively. The protein was dissolved to a concentration of 0.33 mg / mL in the following buffer: 1 x PBS pH 7.4, and 8.9 mg / mL dipalmitoyl phosphatidylcholine. Blood samples were collected from each mouse at the following instants after the injection: 10 minutes, 4 hours, 24 hours and 48 hours. 66

Plasma concentrations of apolipoprotein A-I and derivatives were determined using an ELISA assay, which comprises the following.

Nunc Immuno PolySorp plates were used, each gel was added to each vial at each addition. MB corresponds to the following buffer composition: 2 mM CaCl 2, 1 mM MgCl 2, 10 mM HEPES, 140 mM NaCl.

The plates were coated overnight and in a cooled room with 4 Âμg / ml polyclonal rabbit anti-human apo A-I (DAKO A / S) dissolved in 50 mM NaHC03, pH 9.6. The plates were washed with MB + 0.05% Tween-20, pH 7.8, and blocked with MB + 0.05% Tween-20 + 1% BSA, pH 7.8, for 1 hour. The dissolved sample was applied in MB + 0.05% Tween-20 + 1% BSA, pH 7.8, incubated for 1 hour, washed with MB + 0.05% Tween-20 + 1% BSA, pH 7.8, and incubated for 1 hour with mouse monoclonal anti-apo AI (Perimmune Inc, clone 10-A8) at a concentration of 1 Âμg / ml in MB + 0, 05% Tween-20 + 1% BSA, pH 7.8. The plates were washed and incubated with a secondary anti-mouse IgG antibody linked to horseradish peroxidase. The plates were washed again and allowed to develop using OPD and H202 tablets, the reaction was quenched using H2 SO4 and the result was compared to a standard result based on known concentrations of apo AI and derivatives, respectively. No effect of the dilution of Apo A-I on mouse plasma was observed compared to the standard.

In the results, shown in Figure 14, it is possible to verify that the disappearance time of the Apo A-I Trip A construct increased in the plasma at least 3 times in 67

compared to the disappearance time of native Apo AI. Preliminary data indicate that the disappearance time for Trip A FN Apo A-I is at least identical to Trip A Apo A-I. These data together with the cubilin binding data and the DMPC binding data allow one to conclude that the constructs according to the invention are strong candidates for the treatment of the diseases referred to herein.

Example 9: Plasmids The construct according to the invention can be produced using the following plasmids.

Introduction of a linker sequence in Trip-A-AI

The constructs containing the basic linker, with the referred mutations, were constructed as a construct without the linker. That is, by amplification of Apo A-I (and the linker sequence) and insertion into the plasmid pT7FxH6-Trip-A. The reverse primer was the same as that used for constructing pT7H6FxTrip-A-AI, whereas the forward primers used were: pT7H6FX-Trip-A-FN (-2) -AI: 5'-CGC GGATCC TCG GGT CAG GAT GAA CCC CCC CAG AGC CCC-3 '; unfortunately all isolated clones had the underlined G mutated to a T, indicating an imperfect sequence of the primer.

pT7H6FX-Trip-A-TN-AI-Bam-S 5'-cgc gga tcc aag gtg cac atg aag gat gaa ccc ccc cag age ccc-3 '68

The above mutations were corrected by site-directed mutagenesis using the Stratagene 'QuickChange' kit and the following sets of primers: pT7H6FX-Trip-FN-AI: 5'-acg gtc tg gtg gag acc ggt gag gat g- 3 '5'-cat cct gac ccg agg ttc cct tca ggg aga ccg t-3' pT7H6FX-Trip-A-TN-AI: 5'-acg gtc tg cg ag gag acc aag gtg cac atg aag g-3 '5 '-cct tca tgt gca cct tgg ttc cct tca ggg aga ccg t-3'

Removing the Trip-A Heparin Binding Site

As another variant of the constructs, the heparin binding site of the Trip-A sequence (Lorentsen RH, Graversen JH, et al., Biochemical Journal (2000), 347 pp. 83-87) was mutated using the targeted mutagenesis kit to the site and using the following set of primers: for the lysine 9 mutation from Trip-A: 5'-cca acc cag aga ccc aag gcat aat gta-3 '5'-gtg ttc gca ttt aca atc-3 'to the lysine 15 mutation from Trip-A: 5'-ggc att tac aat cgc ctt ggg ctt gt ggt tgg-3' 5'-cca acc cag aag cg aag gcg att gta aat gcc-3 '

These mutations were planned to be performed on all relevant Trip-A-apo-Al derivatives, and possibly all. Double K9A, K15A mutants of the trip-A derivatives, designated TripA-FN-AI-K9AK15A, TRIP-TN-AI-K9AK15A and TripA-AI-K9AK15A were generated. 69

In addition, truncation of the N-terminus can also remove the affinity for heparin without removing the trimerization. See Holtet et al. Protein Science (1997), Lorentsen et al. Biochem. Journal (2000), Nielsen et al. FEBS (1997) and Nielsen et al. Acta Cryst D. (2000). The N-terminal residue would then preferably be located between Val 16 and Met 22.

Construction of the expression plasmid for Hp-α-A-I

First, the plasmid pT7H6Fx-Hp (a) (Figure 10H) was constructed as follows. From a cDNA library (Clontech, fetal liver) the Hp-α sequence was amplified by PCR using the following set of primers:

Non-sense primer: 5'-cac aag ctt tcc gct aga tct ctg cac tgg gtt age cgg att ctt ggg -3 '; sense primer: 5'-ggt gga tcc ate gag ggt agg ggt gtg gac tea ggc aat gat gtc acg g-3 '. The PCR product had the following characteristics:

Local BamH I- sequence Hp α - local Bgl II - local Hind III.

Following Hp-α cysteine, which forms disulfide bonds with the β chain of Hp, has been mutated to an alanine.

The product was digested with BamHI and Hind III and inserted into plasmid pT7H6FX, which determined the sequence.

A PCR product encoding human Apo A1 was prepared using the standard non-sense primer, which was also used for the preparation of pT7H6-Ubi-Fx-ApoAI and pT7H6Fx-TripA-Al. Whereas the sense primer used was that used as the sense primer for the construction of pT7H6FX-TripA-A1. A PCR product with the following characteristics was obtained: BamH I site - Apo AI sequence - Hind III site, this product was digested with BamHI and Hind III and inserted into the above-mentioned plasmid, digested with Bgl II and Hind III . The resulting pT7H6FX-Hp (a) -Apo AI plasmid was sequenced and the expression in E. coli was tested. pT7H6 TripA-apoAI (Figure 7) The plasmid comprises plasmid pT7H6FxtripA described in example 1 of WO 98/56906. Expression is controlled by the T7 promoter. In addition, the plasmid comprises an H6 sequence, which is a hexa-His affinity tag, for use in the purification. Then a Factor Xa recognition sequence (IQGR) is inserted. SPGT is a connection sequence to the subsequent trimerization module. This sequence has been inserted since it provides the opportunity to cut off the DNA strain with Bgl II and Κρη I. Trip A is the trimerization modulus of tetranectin. GS is another connection sequence, which provides the opportunity to cut with Bam HI.

Finally, the plasmid comprises human apolipoprotein A-1 cDNA encoding amino acids 25 to 267 of apolipoprotein A-I. The expressed and purified protein corresponds to SEQ ID NO 3. 71 pT7H6TripA-apoA-del43 (FIG. 8) The plasmid comprises the above sequences, although the apolipoprotein part is replaced by cDNA encoding amino acids 68 to 267 of human apolipoprotein A-1. The expressed and purified protein corresponds to SEQ ID NO 4. pT7H6UbiFxApoAI (Figure 5). The basic plasmid has been described by Ellgaard et al (1997). The plasmid comprises the following sequences: the expression is controlled by the T7 promoter H6: hexa-His affinity tag for purification of the Ubi: cDNA construct encoding human ubiquitin which has been inserted to stabilize the protein in E. coli-FX: recognition sequence for Factor Xa DNA encoding two Gly residues, which are required for optimal cleavage by Factor Xa-ApoAI: cDNA encoding amino acids 25 to 267 of human apolipoprotein AI.

The expressed and purified protein corresponds to SEQ ID NO 1. pT7H6UbiFXCysApoAI (Figure 6)

As before, but following the sequence encoding the two glycine residues and prior to insertion of the apolipoprotein A-I sequence encoding a cysteine residue. Expressed and purified protein corresponds to SEQ ID NO 2. The plasmid comprises the following sequences: expression is controlled by the T7 promoter H6: hexa-His affinity tag for purification of the Ubi protein construct : CDNA encoding human ubiquitin, which has been inserted to stabilize the protein in E. coli-FX: recognition sequence for Factor Xa DNA encoding two Gly residues, which are required for optimal cleavage by Factor Xa-DNA encoding a cysteine residue - ApoAI: cDNA encoding amino acids 25 to 267 of human apolipoprotein AI.

The expressed and purified protein corresponds to SEQ ID NO 2.

Other examples of plasmids for the expression of apolipoprotein constructs according to the invention are shown in Figures 10A to G together with the corresponding amino acid sequences of the expressed and purified proteins which are described in the sequence listing.

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Neame, P.J. and Boynton, R.E. (1996). Protein Soc. Symposium, (Meeting date 1995; 9th Meeting: Tech. Prot. Chem VII). Proceedings pp. 401-407 (Ed., Marshak, D. R., Publisher:

Academic, San Diego, Calif.).

Nielsen, B.B. (1996). Lecture, Lundbeck Center Neuro-Medicinal Chemistry Minisymposium held November 5, 1996 at the Royal Danish School of Pharmacy, Copenhagen.

Nielsen, B.B., Larsen, I.K., Rasmussen, H. and Kastrup, J.S. (1996). Lecture, Danish Crystallographer's Meeting, held June 3-4, 1996 at the Royal Danish School of Pharmacy,

Copenhagen.

Siebwenlist et al. 1980. Cell, 20: 269. Surgensen et al., 1995, Gene, 152: 243-245. Stinchomb et al. 1979. Nature 282: 39. Tschemper et al. 1980. Gene, 10: 157. 75

SEQUENCE LISTING < 110 > Proteopharma ApS < 120 > Apolipoprotein Analog < 130 > P 459 PC00 < 150 > DK PA 2000 01682 < 151 > 2000-11-10 < 150 > DK PA 2001 00057 < 151 > 2001-01-15 < 150 > US 60/264, 022 < 151 > 2001-01-26 < 160 > 14 < 170 > Patentln version 3.1

< 210 > 1 < 211 > 243 < 212 > PRT < 213 > Homo sapiens < 400 > 1 76

Asp Glu Pro Pro Gin Ser Firo Trp 1 S

Go Tyr Go Asp Go Leu Lys Asp 20

Fhe Glu Gly Ser Ala Leu Gly Lys 35 40

Asa Trp Mp Ser Vai Thr Ser Thr 50 55

Gly Pro Goes Tbr Gin. Glu Phe Trp

Gly Leu Atg Gla Glu Met Ser Lys SS

Go Gin Gin Tyr Leu Asp Asp Phe 100

Gin Leu Tyr Arg Gin Lys Tal Glu 115 120 ASp Arg Vai Lys Asp Leu Ala Thr 10 15 Ser Gly Arg Asp Tyr Will Be Gin 2S 30 ei »Leu Asa Leu Lys Leu Leu Asp 45« Ha be Lys Leu «Arg Glu Gin Leu Aep Asn Leu Glu Lys Glu Thr Glu 75 80 Asp Leu Glu Glu Go Lys Ala Lys SO 95 Gin Lyn Lys Trp Gin Glu Glu Met 105 110 Pro Leu Arg Ala Glu Leu Gin Glu 125 77

Gly Ala Arg Gin Lys Leu. Hís Glu Leu Gin Gin. Lys Leu ser pro leu 130 135 IIP

Gly Olu Glu Met Arg Asp Arg Ala, Arg Ala Mia Vai Asp Ala Leu Arg MB ISO 155 160

Thr Sis tu Ala Pro fyr Ser Asp Glu Leu Arg Gin Arg Leu Ala Ala 165 170 175

Arg Leu Glu Ala Leu Lys Glu Asu Gly Gly Ala Arg Leu Ala Glu fyr ISO 185 196

Ala Lys Ala Thr Glu Sis Leu Ser ffar Leu being Glu Lys Ala Lys 195 200 205

Pro Ala Leu Glu Asp Leu Arg Gin Gly Leu Leu Pro Vai Leu Glu Ser 310 215 220

Phe Lys will be Phe Leu Ser Ala Leu Glu Glu Tyr TAr Lys Lys Leu 22S '230 235 240 Ãsn Thr Glu

&lt; 210 &gt; 2 &lt; 211 &gt; 244 &lt; 212 &gt; PRT &lt; 213 &gt; Homo sapiens &lt; 220 &gt; &lt; 221 &gt; MI ST _FEATURE &lt; 222 &gt; (244) <223> Amino Acids 25 to 267 of the compound of the formula: ## STR1 ## in which: Human Apo Al <400 &gt; 2

Cys Asp Glu Pro Pr o Gin Ser Pro pSe pg Go to Asp Leu. Ik l '5 10 15

Thr Will Tyr Will Asp Will Asp Asp Ser Gly Arg Asp Tyr Will Be 50 25 30

Gin Ph§ Glu Gly Ser ala Leu Gly Lys Gin Leu Asn Leu Lys Leu Leu 79 35 40 45

Asp Mn Trp Mp Ser will Thr be Thr Fite Ser Lys thy Arg Glu Gin 50 55 60

Leu Gly Fro goes Tbr Gin Glu Fhe Trp Asp Asa Leu Glu Lys Glu Thr 65 70 75 SO

Glu Gly Leu & Gla GluMet Ser Lys Asp Leu Glu Glu Val Lys Ala 55 90 95

Lys Go Gin Fro Tyr Leu Asp Asp Phe Gin Lys Lys Trp Glu Glu Glu 100 105 110

Net Glu Leu Tyr Arg Gin Lys Go Glu Pr © Leu Arg Ala Glu Leu Glu 115 120 125

Glu Glv Ala Arg Gin Lys Leu Bis Glu Leu Gin Glu Lys Leu Ser Fro ISO 135 U0

Leu Gly Glu Glu Met Arg Asp Arg Ala Arg Ala His vai Asp Ala Leu 145 150 155 ISO

Arg Tbr His Leu Ale Fro Tyr Ser Asp Glu Leu Arg Gin Arg Leu Ala 165 170 175

Ala Arg Leu Glu Ala Leu Lys Glu Asn Gly Gly Ala Arg Leu Ma Glu ISO 185 190

Tyr His Ala Lys Ala Tbr Glu Bis Leu Ser Thr Leu Ser Glu Lys Ala 195 700 205

Lys Fro Ala Leu Glu Asp Leu Arg Glu Gly Leu Leu Fro vai Leu Glu 210 215 220

Lys will be Pbe Leu be Ala Leu Glu Glu Tyr Tbr Lys Lys 225 230 235 240

Leu Asn Thr Glu, 80

&lt; 210 &gt; 3 &lt; 211 &gt; 301 &lt; 212 &gt; PRT &lt; 213 &gt; Homo sapiens &lt; 220 &gt; &lt; 221 &gt; misc_feature &lt; 222 &gt; (1) .. (58) &lt; 223 &gt; Trimerization module from tetranectin &lt; 220 &gt; &lt; 221 &gt; MISC_FEATURE &lt; 222 &gt; (59). . (301) &lt; 223 &gt; Apo Al mature &lt; 400 &gt; 3 81 be Pro Gly Thr Glu Pro Pro Thr 01a Lys Pro Lys Lys Ile Val Asa 1 5 10 15

Ala Lys Lys Asp will Va Asa Thr Lys mt Phe Glu Glu hm Lys Ser 20 2S 10

Ares Leu Asp Thr Leu Ala Gin Glu Vai Ala Leu Leu Lye Glu Gin Gin 35 40 45

Ala Leu © Thr Will Be Leu Lys Gly Ser Asp Glu Pro Pro Gin Ser S0 55 60

Pro Trp Asp Ara Vai Lys Asp Leu Ala Thr Vai Τγ £ 'Vâl Asp Vai Leu 65 70? S 80

Lys Asp Ser Gly Arg Asp Tyr Will Be Gin Phe Glu Gly Ser Ala Leu 65 90 25

Gly Lys Gin Leu Asa Leu Lys Leu Leu Asp Asn Trp Asp Ser Vai Thr 100 105 110

Ser Tbr Phe Ser Lys Leu Arg Glu Glu Leu Gly Pro Val Tbr Gin Glu 115 120 125

Fhe Trp Asp Asn Leu Glu Lys Glu Thr Glu Gly Leu Arg Glu Glu Ffet 130 13S 140

Ser Lys Asp Leu Glu Glu Go Lys Ala Lys goes Gin Pro Tyr Leu Asp 145 ISO 1SS 160

Asp 1% and Gin Lys Lys Trp Gin Glu Glu Met Glu Leu Tyr Arg Gin Lys 165 1? 175

Go Glu Pro Leu Arg Ala Glu Leu Glu Glu. Gly Ala arg Gin Lys Leu 180 185 120 82

His Glu Leu Glu Glu Lys Leu Ser Pro Leu Gly Glu Glu Sfet Arg Asp 195 200 2SS

Arg Ala Arg Ala His Va Asp Ala hm Arg Thr His hm Ala Pro Tyr 210 215 220

Ser Asp Glu Leu Arg Gin Arg Leu Ale Ma Arg Leu Glu Ala Leu Lys

225 230 235 24Q

Glu A9U Gly Gly Ma Arg Leu Glu Glu Tyr His Ala Lys Ala Thr Glu

245 Ξ50 2SS

His Leu Ser Thr Leu Ser Glu Lys Ma Lys P.ro Ala Leu Glu Asp Leu 260 ZeS 270

Arg Gin Gly Leu Leu Pro Go Leu Glu Be Phe Lys Will Be Phe Leu 275 283 285

Be Lea Wing. Glu Glu Tyr Thr Lys Lys Leu Asn Thr Glu 200 2§S 330

&lt; 210 &gt; 4 &lt; 211 &gt; 258 &lt; 212 &gt; PRT &lt; 213 &gt; Homo sapiens &lt; 220 &gt; &lt; 221 &gt; MISC_FEATURE &lt; 222 &gt; (1) . . (58) &lt; 223 &gt; Trimerization module from tetranectin &lt; 220 &gt; &lt; 221 &gt; MISC_FEATURE &lt; 222 &gt; (59) .. (258) &lt; 223 &gt; Amino acids Apo-68-267 human &lt; 400 &gt; 4 Serr. Oly Thr., Pro Pr. Thr. Lya Level Asn 1 5 10 15 M & Lvs hy® Asp Vâl Vel Asn Thr hys and Bhe Glu (31u ItfiU Lys be 30 25 30 Arg Asp Thr heii Ala Gin 01 a Vai Alâ IiOU Leu Lys Glu oia Gin 35 40 45 84

Wing Leu Gin Tbr Will Be Leu Lys Gly Be Leu Lys Leu Leu Asp Asn. only so

Trp Asp Ser Thi Be Ser Thr Fhe Be Lys Leu Arg Glu Glu Leu Gly 63 70 75 80 *

Pr © Go Thr Gin Glm Ptoe Trp Asp Asn Leu Glu Lys Glu Thr Glu Gly 85 90 95

Leu Arg Gin Glu Met Ser Lys Asp Leu Glu Glu goes Lys Ala Lys goes 10 0 103 no

Glu Pr o Tyr Leu Asp Asp Phs Gin Lys Lys Trp Gin Glu Glu Het Glu 115 120 12S

Leu Tyr Arg Gin Lys Go Glu Pro Leu Arg Ala Glu Leu Glu Glu Gly 130 135 140

Ala Arg Gin Lye Leu Kie Glu Leu Glu Glu Lys Leu Ser Pro Leu Gly 145 250 155 160

Glu Glu Met Arg Asp Arg Ala Arg Ala Ris Go Asp Ala Leu Arg Thr 1IS 170 175

Ris Leu Ala Pro Tyr Ser Asp Glu Leu Arg Gls Arg Leu Ala Ala Arg 180 185 19

Leu Glu Alâ Leu Lys Glu Asn Gly Gly Alâ. Arg Leu Ala Glu Tyr Sis 195 200 205

Ma Lys Ma Thr Glu Sis Leu Ser Thr Leu Ser Glu Lys Ala Lys Pro 210 215 220

Ala Leu Glu Asp Leu Arg Gin Gly Leu Leu pro Vai Leu Glu Ser Phe 225 230 235 240

Lys Will Be Fhe Leu Be Lea Leu Glu Glu Tyr Thr Lys Lys Leu Asn 245 250 255

Thr Gin 85

&lt; 210 &gt; 5 &lt; 211 &gt; 301 &lt; 212 &gt; PRT &lt; 213 &gt; Homo sapiens &lt; 220 &gt; &lt; 221 &gt; MISC_FEATURE &lt; 222 &gt; (1) .. (58) &lt; 223 &gt; Trimerization module from tetranectin &lt; 220 &gt; &lt; 221 &gt; MUTAGEN &lt; 222 &gt; (9) .. (9) &lt; 22 3 &gt; &lt; 220 &gt; &lt; 221 &gt; MUTAGEN &lt; 222 &gt; (15) .. (15) &lt; 22 3 &gt; &lt; 220 &gt; &lt; 221 &gt; MISC _FEATURE &lt; 222 &gt; (59) .. (301) &lt; 2 2 3 &gt; Mature Apo-AI &lt; 400 &gt; 5 86

Ser Pro Gly Thr Glu Pro pro Thr Gin Lys Pm Lys Ala ile tal Asa i 15

Wing Lys Wing Asp Will Go Asn Thr Lys Met Phe Gla Qau Mu Lys Ser 20 2 $ 30

Arg Leu tep Thr Leu Ala Gla olu Go .Ala Leu Leu Ly »Glu Gin Gin 35 40 45

Ala Leu Gla Thr Will Be Leu Lys Gly Ser Asp Glu Pro Pro Gla Ser m 60

Pro Trp Asp Arg Go Lys Asp Leu Ala Thr Go Tyr Go Asp Go Leu 65 70 75 SO

Lys Asp Ser Gly Arg Asp Xyr Will Be Gin Phe Slu Gly Ser Ala Leu δS, 90 secs

Lys Gin Leu Mn Leu Lys Leu Leu Asp · Asn Trp Asp Ser Vai Thr 100 105, 110

Ser Thr Phe Ser Lys Leu Arg Glu Gin Leu Gly Pro Go Thr Gla. Glu 115 130 125 87

Phê Trp Aap Asií Leu Glu Lys Glu Thr Glu Gly Leu Arg Gin Glu Met 130 13 S

Ser Lys Asp Leu Glu Glu Val. Lys lala Lys Go Glu Pro Tyr Leu Asp L4§ 150 155 ISO AEp Fha Gin Lys Lys Trp Gin Glu Glu Met Glu Leu Tyr Arg Glu Lys 165 170 175

Go Glu Pro Leu Arg Ala Glu Leu Gin Glu Gly Ala Arg Gin. Lys Leu 180 185 190

This is Glu Leu Glu Glu. Lys Leu Ser Fro Leu Gly Glu Glu Mefc Arg Asp 155 200 205

Arg Ala Arg Wing Ala Leu Arg Thr His Leu Ala Pro Tyr 310 215 220

Ser Asp Glu Leu Arg Gla Arg Leu Ala Arg 'Leu Glu Ala Leu Lys 225 230 235

Glu Asa5Gly Gly Ala Arg Leu. Ala Glu Tyr Bis Ala Lys Ala Thr Glu 245 250 2.55

His Leu Ser Thx Leu Ser Glu Lys Ala Lys Pro Ala Leu Glu Asp Leu 260 265 270

Lys Will Be Phe Leu 2S.S

Arg Gin Gly Leu. Leu Pro Vai Leu Glu Ser Fha 275 280

Ser Wing Leu Glu Glu Tyr Thr Lys Lys Leu Mn Thr Gin 230 295 '300 &lt; 210 &gt; 6 &lt; 211 &gt; 304

&lt; 212 &gt; PRT &lt; 213 &gt; Homo sapiens &lt; 220 &gt; &lt; 221 &gt; MISC_FEATURE &lt; 222 &gt; (1) .. (58) &lt; 223 &gt; Trimerization module from tetranectin &lt; 220 &gt; &lt; 221 &gt; MISC_FEATURE &lt; 222 &gt; (59). . (61) &lt; 223 &gt; Linker &lt; 220 &gt; &lt; 221 &gt; MISC_FEATURE &lt; 222 &gt; (62). . (304) &lt; 223 &gt; Mature Apo-AI &lt; 400 &gt; 6 89

Ser Pro Gly Thr Glu Pro Pro Thr Gin Lys Pro Lys Lys

Wing Lys Lys Mp Will Go Asn Thr Lys Hat Phe Glu Glu Leu. Lys Ser 20 25 30

Arg Leu Asp Thr Leu Ala Gin Glu will Ala Leu Leu Lys Glu Gin Gin 35 40 45

Wing Leu Gin Thr Will Be Leu Lys Gly Be Ser Gly Be Asp Glu Pro 50 55 60

Pro Gin Ser Pro Trp Asp Arg Go Lys Asp Leu Ala Thr Go Xyr Go Go 65 70 75 80

Asp VlL Leu Lys Asp Ser Gly Arg Asp Tyr Will Be Glu Phe Glu Gly 85 90 95

Be Ala Leu Gly Lys Gin Leu Asn Leu Lys Leu Leu Asp Asa Trp Asp 1.00 105 110

Ser Yal Thr Ser Thr Fhe Ser Lys Leu Arg Glu Gin Leu Gly Pro Val 115 115 125

Thr Glu Glu Pfee Trp Asp Asn Leu Glu Lys Glu Thr Glu Gly Leu Arg 230 135 MS

Gin Glu Met Ser Lys Asp Leu Glu Glu will Lys Ala Lys Go Gin. Pro 145 25 è 155 160

Tyr Leu Asp Asp The Gin Lys Lys Trp Gin Glu Glu Met Glu Leu Tyr 165 '170 · 175

Arg Gin Lys Go Glu Pro Leu Arg Ala Glu Leu Gin Glu Gly Ala Arg ISO 185 190

Gin Lys Leu His Glu Leu Gin Glu Lys Leu Ser Pro Leu Gly Glu Glu 195 200 205 90 Mfcg Arg Arg Alkaline Aep Ala Leu &amp; Thr His Leu 210 315 220

Ala Pro Tyr Ssr Asp Glu Mm Arg Gin Arg Leu Ala Ala Arg leu Glu 225 230 23S 240

Wing Thousand Lye Glu Ma Gly Gly Ala Arg Leu Wing <3.1 u Tyy His Wing Lys 245 2S0 25 $

Ala Thr Glu His Leu Ser Thr Leu Ser Glu Lys Ala Lys Pro Ala Leu 260 2S 270

Slu Asp Leu Arg Glu Gly Leu Mu Pro Leu Glu Ser Pbe Lys Go 275 280 28S

Ser Pie Leu Ser Ala Leu Glu Glu Tyr Thr Lys Lys Leu Asn Thr Gin 2S0 2B5 3O0

&lt; 210 &gt; 7 &lt; 211 &gt; 304 &lt; 212 &gt; PRT &lt; 213 &gt; Homo sapiens &lt; 220 &gt; &lt; 221 &gt; MISC_FEATURE &lt; 222 &gt; (1) . . (56) &lt; 223 &gt; Trimerization module from tetranectin &lt; 220 &gt; &lt; 221 &gt; MISC_FEATURE &lt; 222 &gt; (57) .. (61) &lt; 223 &gt; Fibronectin-based linker &lt; 220 &gt; &lt; 221 &gt; MISC_FEATURE &lt; 222 &gt; (62). . (304) &lt; 223 &gt; Αρο-ΑΙ mature &lt; 400 &gt; 7

Be Pro ôly Thr Glu Pro Pro Tk Glrs Lys Pro Lys Lys He Go Mn XO 15

Ma Lys Lys Mp Will Go To Thr Lys Met Phe Glu Glu Leu. Lys Ser 20 25 30

Arg Leu Âsp Thr Léu Ma 01 Â »Glu Vai Ala Leu Leu Lys Glu Oln Glu 3S 40 45

Ser Phe leu Ser Wing Leu Glu Glu Tyr Thr Lys Lys Leu Asn Thr Gin 290 2P5 300

&lt; 210 &gt; 8 &lt; 211 &gt; 304 &lt; 212 &gt; PRT &lt; 213 &gt; Homo sapiens &lt; 220 &gt; &lt; 221 &gt; MISC_FEATURE &lt; 222 &gt; (1) .. (56) &lt; 223 &gt; Trimerization module from tetranectin &lt; 220 &gt; &lt; 221 &gt; MISC_FEATURE &lt; 222 &gt; (57) .. (61) 92 &lt; 223 &gt; Frieronectin-based linker &lt; 220 &gt; &lt; 221 &gt; MISC_FEATURE &lt; 222 &gt; (62). . (304) &lt; 223 &gt; Mature Apo-AI &lt; 22 0 &gt; &lt; 221 &gt; MUTAGEN &lt; 222 &gt; (13) .. (13) &lt; 22 3 &gt; &lt; 22 &gt; &lt; 221 &gt; MUTAGEN &lt; 222 &gt; (19) .. (19) &lt; 22 3 &gt; &lt; 4 0 0 &gt; 8

Ser Pro Gly Tfir 01¾ Proferro Tfer GXn Lys Pro Lys Ala rie Goa Asa 1 5 10 is

Ma Lys Ala Asp Go. Go Asa Tbr Lys Met Pb® Gin Glvt Leu Lys Ser 20 25 30

Arg Leu. top Thr Leu Ala Gin Slu Vai Ala Leu. Leu Lys Glu Gin Gin 3S 40 4S

Ala Leu Ola Thr Will Be Leu Lys Gly Tbr Be Gly Gin Asp Glu PrO B0 3S SQ

Pro Gin Ser Pro Trp Asp Arg Go Lys Asp hm Ais Thr Go Tyr Go 93 «S 70 75 80

Asp will Leu Lys Asp Ser Gly Arg Asp Tyr Will Be Gin Phe Glu Gly 85 80 85

Ser &amp; Leu Gly Lyg Qln lsu Asa Leu Ly $ Leu Leu, Asp Asa Trp Asp 100 105 110

Ser Thy Sar Thr Thy Ser Lys Leu Arg Glu Gls Leu Gly Pro Val 115 115 125

Thr Gltt Glu Phs Trp Asp Asn Leu Glu Lys Glu Thr Glu Gly 'Leu Ara 130 135 140

Gin Glu Met Ser Lys Asp Lsu Glu Glu Val Lys Ala Lys. Vai Gla Pró 145 ISO 155 ISO

Tyr Lsu Asp Asp Phe Gin Lys Lys Trp Gin Glu Glu Met Glu Leu Tyr 1S5 170 175

Arq Gla Lys Go Glu Pr Leu & Rq Ala Glu Leu Gin Glu Gly Ala Arg ISO 1S5 180

Gin Lys Leu His Glu Leu Gin Glu Lys Lsu Ser Pro Leu Gly Glu Glu 135 300 205

Met Arg Asp Arg Ala Arg Ala His Vai Asp Ala Leu Arg Tfer llis Leu 210 215 220

Ala Pro Tyr Ser Asp Glu Leu Arg Gin Arg Leu Ala Ala Arg Leu Glu 225 '23S 235 240

Ala Leu Lys Glu Asa Gly Gly Ala Arg Leu Ala Gla Tyr Eis Aja Lys 245 250 255

Ala Thr Glu His Leu Ser Thr Leu being Glu Lys Ala Lys Pro Ala Leu 280 265 270

Glu Asp Leu Arg Glu Gly Leu Leu Pro Go Leu Glu Ser Phe Lys Go 275 280 285

Ser Phe Leu Ser Ala Leu Glu Glu Tyr Thr Lys Lys Leu Asn Thr Glu 2SQ 295 300 94 &lt; 210 &gt; 9 &lt; 211 &gt; 306 &lt; 212 &gt; PRT &lt; 213 &gt; Homo &lt; 22 &gt; &lt; 221 &gt; MISC_FEATURE &lt; 222 &gt; (1) .. (58) &lt; 223 &gt; Trimerization module from tetranectin &lt; 220 &gt; &lt; 221 &gt; MISC_FEATURE &lt; 222 &gt; (59) .. (63) &lt; 22 3 &gt; Linker &lt; 220 &gt; &lt; 221 &gt; MISC_FEATURE &lt; 222 &gt; (64) .. (606) &lt; 223 &gt; Mature Apo-AI &lt; 400 &gt; 9 95

be Pro Gly Thr Glu Pro Pro Thr Gin Lys Pro Lys Lyg ile Go Aan X 5 10 IS

Ala Lys Lys Asp Go Go Asa Thr Lys Mefc Phe Glu Glu Leu Lys Ser 20 25 30 Arg Leu Asp Thr Leu Ma she she Go Ala Leu. Leu Lys Glu Gin Gin. 35 40 45

Wing Leu Gin Thr Will Be Leu Lys Gly Be Lye Will His Met Lys Asp 50 55 S0

Glu Pro Pro Gin Ser Pro Trp Asp Ârg Go Lys Asp Leu Ala Thr Go € 5 70 75 00

Tyz Go Asp Go Leu Lys Asp Ser Gly Arg Asp Tyr Will Be Gin Phe 85 50 95

Glu Gly Ser Ala Leu Gly Lys Gin Leu Asn Leu Lys Leu Leu Asp Asn 100 X05 110

Trp Asp Ser Thr Thr Ser Thr Phe Ser Lys Leu Arg Glu Gin Leu Gly 115 120 125

Pm Go Thr Gin Glu Phe Trp Asp Asn Leu Glu Lys Glu Thr Slu Gly 130 135 140 96 Leu Arg 31a Glu Met Ser Lys 145 ISO Glu Pro Tyr Leu Asp Asp Fhe 165 Leu Tyr Arg Olu Lys Go Glu ISO Ala Arg Glu Lys Leu His Glu 195 Glu Glu Mefc Arg Asp Arg Ala 210 215a Leu Ma Pro Tyr Ser Asp 225 230 Leu Glu Ala Leu Lys Glu Asu 245

Glu Glu Go Lys Ala Lys Go 155 * 160

Lys Itys Trp δία Glu Glu Met Glu 170 175 leu Arg Ma Glu Leu Gla Glu Gly 165 190

Gin Glu Xtvs Read Be Pro Your Gly 205

Ala His Vai Asp Ala His Arg Thr 220

Your Arg Arg Arg Your Ala Arg Arg 235 240

Gly Ala Arg His Glu Tyr His Wing 2SQ 255

Ala Lys Ala Thr Glu Eis Leu 26 ©

Thr Your Ser Glu Lys Ma Lys Pro 265 270

Ala leu Asp hm Arg om 275

Read Your Fm vm Leu Glu be Phe 285

Lys Will Be Phe Leu Ser Ma 290 295

Glu Glu Tyr Thr Lys Lys Leu Asn 300

Thr Gin

3 · OS &lt; 210 &gt; 10 &lt; 211 &gt; 306

&lt; 212 &gt; PRT 97 &lt; 213 &gt; Homo sapiens &lt; 220 &gt; &lt; 221 &gt; MISC_FEATURE &lt; 222 &gt; (1) .. (56) &lt; 223 &gt; Trimerization module from tetranectin &lt; 220 &gt; &lt; 221 &gt; MISC_FEATURE &lt; 222 &gt; (57) .. (63) &lt; 223 &gt; Linker based on tetranectin &lt; 220 &gt; &lt; 221 &gt; MISC_FEATURE &lt; 222 &gt; (64) .. (306) &lt; 223 &gt; Mature Apo-AI &lt; 400 &gt; 10 98

Be Pro Gly Thr Glu Pro pro Thr Qla Lys P £ * Lys Lys IXe Go Asa 1 5 10 15

Ala Lys Lys Asp Go. Go Asn Thr Lys Mefe PM Glu Gin Leu Lys Ser ae as 30

Arg Leu Asp Thr Leu Ala Gin Glu Go Ala hm Leu Lys Glu Gin Gin SS 40 45

Ala Leu Glu Thr will be Leu Lys Gly Thr Lys Willi SSet Lys Aep

Glu Ρϊώ Pro Gin Ser Pr * Trp AS »Afg vai Lys Asp Leu Ma Thr Go 65 70 75 SG

Tyr Go Aöp Go Leu Lys Ásp Ser Gly Arg Mp Tyr Gonà © Ser Gin Phe 85 50 55

Glu Gly Ser Ala Leu Gly Lys Gin Leu Asa Leu Lys Leu Leu Asp Ãen 100 105 110

Tip Asp Ser Go Thr Ser Thr Phe Sei- Lys Leu Arg Glu Glu Leu Gly 115 120 125

Pro Go Thr Gin Glu PM Trp Mp Wing Leu Glu Lys Glu Thr Glu Gly 130 135 140

Leu Arg Gin Gin Mefc Ser Lys Asp Leu Glu Glu Go Lys Ma Lys Go 14S 150 155 ISO

Gin Pro Tyr Leu Asp Asp Phe Gin Lys Lys Trp Gin Glu Gin Met Glu 155 170 175

Leu Tyr Arg Gin Lys Go Glu Pro Leu Arg Wing Glu Leu Gin Glu Gly 180 185 130

Ala Arg Glu Lys Leu His Glu Leu Glu Gin Lys Leu Ser Pro Leu Gly 155 200 205 99

Glu-Glu-Arg-Arg Asp-Arg-Arg-Arg-Ala-Hls-Asp-Ala-Leu-Arg-

Eis Leu Ala Pro Tyx Ser &amp; Glu Leu Arg Gin Arg Leu Ala Ala Arg 225 230 235 240

Leu Glu Wing Leu Lys Glu Wing. Gly &lt; &amp; Ala Arg Lêu Ala Glu Tyr Eis

245 2S0 25S

Ma. Lys Ala Thr Glu Hís thy Thr Leu Ser Glu Lys Ala Lys Pro 2m 265 270

Ala Leu Glu Aap Leu Arg Gin Gly Leu Leu Pro Go Leu Glu Ser Phe 27S * 289 285

Lys Will Be Phe Leu Be Wing Leu Glu Glu Tyr Thr Lys Lys Leu Asn

220 2 $ S MG

Thr Gin 305

&lt; 210 &gt; 11 &lt; 211 &gt; 306 &lt; 212 &gt; PRT &lt; 213 &gt; Homo sapiens &lt; 220 &gt; &lt; 221 &gt; MISC_FEATURE &lt; 222 &gt; (1) .. (56) &lt; 223 &gt; Trimerization module from tetranectin &lt; 220 &gt; &lt; 221 &gt; MISC_FEATURE &lt; 222 &gt; (57). . (63) 100 &lt; 223 &gt; Linker based on tetranectin &lt; 220 &gt; &lt; 221 &gt; MISC_FEATURE &lt; 222 &gt; (64) .. (306) &lt; 223 &gt; Mature Apo-AI &lt; 220 &gt; &lt; 221 &gt; MUTAGEN &lt; 222 &gt; (13) .. (13) &lt; 2 2 3 &gt; &lt; 220 &gt; &lt; 221 &gt; MUTAGEN &lt; 222 &gt; (19) .. (19) &lt; 2 2 3 &gt; &lt; 400 &gt; 11 101

Ser Pro Gly Thr Glu Pro Pro Thr β, Lys Pro Lys Ala Xis Go Asa as w is

Ma Lys Asp Asp goes Asa Thr Lys Met Phe Glu Glu Leu Lys Ser 20 25 30

Arg Leu Asp Thr Leu Ala Gin Glu Vai Ala Leu Leu Lys Glu Gin Glu 35 40 45

Ala Leu Glu Thr Will Be Leu Lys Qlv Thr Lys Will Sis Met Lys Asp 50 55 S0

Glu Pro Pro Gin Ser Pro Trp Asp Arg Go Lys Asp Leu Ala Thr Go fiS 70 75 00

Tyr Vsi Mp will Leu Lys Mp Be Gly Arg Mp Tyr will be Glu phe 85 90 95

Glu Gly Ser Ala Leu Gly Lys Glu Leu. Asa Leu Lys Leu Leu Asp Asa. 100 105 · 110

Trp Asp Ser Be Thr Ser Thr Phe Ser Lys Leu Arg Glu Gin Leu Gly 115 120 * 125

Pro Go to Thr Gin'Glu Phe Trp Asp hm Leu Glu Lys Glu Thr Glu Gly 130 135 40

Leu Arg Glu Glu Met Ser Lys Asp Leu Glu Glu Val Lys Ala Lys Val 14S 150 155 160

Glu Pro Tyr Leu Asp Asp Phe Glu Lys Lys Trp Gin Glu Glu Met Glu LES 170 175

Lsu Tyr Arg Gin Lys Go Glu Pro Leu Arg -Ala Glu Leu Glu Glu Gly 180 185 190

Ala Arg Gin Lys Leu His Glu Leu Gin Glu Lys Leu Ser Pro Leu Gly 195 200 205

Glu Glu Met Arg Asp Arg Ala Arg Ala Ala Asp Ala Leu Arg Thr 21.0 '215 220 102 ais Leu Ala Pro Tyr be Asp Glu Leu Arg Gin Arg Leu Ala Ala Arg 225 230' 23 S 240

Leu Glu Ala Leu Lys Glu Asn Sly Gly Ala Arg Leu Ala Glu Tyr Kis 245 250 355

Ala Lys Ala Thr Glu His Leu Ser Tke Leu being Glu Lys Ala Lys Pro 260 20Ξ 270

Ala Leu Glu Asp Leu Arg Glu Gly Leu Leu Pro Vai Leu Glu Ser Pha 275 260 285

Lys Will Be Phe Leu Be Wing Leu Glu Glu Tyr Thr Lys Lys Leu Asn SSO 2SS 300

Tftr Gin 305

&lt; 210 &gt; 12 &lt; 211 &gt; 51 &lt; 212 &gt; PRT &lt; 213 &gt; Homo sapiens &lt; 3 0 0 &gt; &lt; 302 &gt; Trimerisation module &lt; 30 &gt; &lt; 310 &gt; WO 98/56906 &lt; 311 &gt; 1998-06-11 &lt; 312 &gt; 1998-12-17 &lt; 313 &gt; (1) . . (51) &lt; 4 0 0 &gt; 12 103

Glu Pro Pro Thr Gin Lys Pro Ly »Lye Ile Vai Mn Ala Lys Lys Asp 1 5 10 15 Val Val Asa Tfcr Ly» mt Phs Glu Glu Leu Lys Se? Arg Mu Asp Thr 20 25 30

Leu Aia Gin Glu Goes Ala Mu Mu Ly * Glu Gl »Si» Ala Mu Gla ffcr 35 40 45

Go Cys Leu

&lt; 210 &gt; 13 &lt; 211 &gt; 58 &lt; 212 &gt; PRT &lt; 213 &gt; Homo sapiens &lt; 220 &gt; &lt; 221 &gt; MISC_FEATURE &lt; 222 &gt; (1) .. (4) &lt; 223 &gt; Linker Sequence &lt; 220 &gt; &lt; 221 &gt; MISC_FEATURE &lt; 222 &gt; (5) .. (56) &lt; 223 &gt; TTSE modified &lt; 2 2 0 &gt; &lt; 221 &gt; MISC_FEATURE &lt; 222 &gt; (57) .. (58) &lt; 2 2 3 &gt; Linker &lt; 4 0 0 &gt; 13 104

Ser Pro Giy Tfer 81 «Pre * Pro Tbr Sln kys Pro Dy» ys lie vai ásn 1 5 10 15

Ala Lys hys Asp Go Va Asa Thr i «» Met Phe <3lu? Lys Ser 30 25 30

Arg. Th.r Leu Ala Gin Slu Goes Ala Leu Leis. Ly &amp; 61 «Gltt 61« 35 40 45

Ala Leu 61b T &amp; R will be Leu Lys Gly Ser 50 53

&lt; 210 &gt; 14 &lt; 211 &gt; 329 &lt; 212 &gt; PRT &lt; 213 &gt; Homo sapiens &lt; 220 &gt; &lt; 221 &gt; MISC_FEATURE &lt; 222 &gt; (1) .. (86) &lt; 223 &gt; Hp (alpha) residues &lt; 220 &gt; &lt; 221 &gt; MISC_FEATURE &lt; 222 &gt; (87) .. (329)

&lt; 223 &gt; Apo A-I &lt; 400 &gt; 14

Gly Go Asp Ser Gly Aa a Asp Go Thr Asp Ile Ala Asp Asp Gly Cys .1 5 10 IS 105

Pro Lys pro Pro Glu Ile ala His Gly Tyr is Glu His ssr Go to asa 20 25 30

Tyr Gin cys Lys wing Tyr Tyr Lys Leu Arg Thr Glu Gly sp siy vai 35 43 4S

Tyr Thr i * eu Asn asa Glu Lys Glu Trp He Ase Lya Ala Go Gly Asp 50 S5 50

Lys Leu Pro Glu Cys Glu AM-V & L Ala Gly Lys Pm Ly »Wing Pm Ala 65? 0 75 00

Asp Pro Val Gin Arg Asp Glu Pro Pro Glu Ser Pro Trp Asp Arg 85 90 95

Val Lys Asp Leu Ala Thr Val Tyr Val Asp Val Leu Lys ASp Ser Gly 103 105 U

Arg Agp Tyr Val Ser Gin Phe Glu Sly Ser Ala Leu Gly Lys Gin Leu 115 120 125

Asn Leu Lys Leu Leu Asp Asp Trp Asp Ser Val Thr Ser Thr Phe Se 133 133 140

Lys Leu Arg Glu Gin Leu Gly Pro Val Thr Gin Glu Pfte Trp Asp AsA 14S 150 15S ISO

Leu Glu Lys Glu. Thr Glu Gly Leu Arg Gin Glu Ket Ser Lys Asp Leu 1 170 170 175

Glu Glu Vai Lys Ala Lys Vsl Gin Pm Τ '/ r Leu & sp Asp Phe Gin Lys 180 185 190

Lys Trp Gin Glu Glu fíet G & U Leu Tyr ..Arg Gin Lys Val Glu Pro Leu 135 200 205

Arg Ala Glu Leu Sln Glu Gly Ala Arg Si »Lys Leu Sis Glu Leu Gin 210 215 220

Glu Lys Leu Ser Pro Lau Gly Glu Glu Mefc Arg Asp Arg Ala Arg Ala 225 230 33S 24α

His Val Asp Ala Lau Arg Thr His Leu Ala pro Tyr be Mp Glu Leu 245 2S0 255 106

Krq Gin Arg kti Ala, Ala Arg Leti Glu Ala Leu Lys Glm Aaa Giy Giy 260 2S5 270

Ala Arg Leu Ala Slu Tyr Bis Ala Lys Alahr Glu His Leu Ser Thr 27S 2-80 285

Leu Ser Glw Lys Ala Lys Pr o Ala Leu, Glu Asp Leu Arg Gin Gly Leu 200 2m 300

Leu Fro Go Leu Glu Be Phe Lys Will Be Phe Leu Be Ml Leu Glu 30S 320 315 320

Glu Tyr Thr Lys Lys Leu Asa Thr Gin 325 SEQUENCE LISTING [0195]

&lt; 110 &gt; Proteopharma ApS &lt; 120 &gt; Apolipoprotein Analogs &lt; 130 &gt; 16278EP00 &lt; 160 &gt; 39 &lt; 170 &gt; Patentln version 3.3

&lt; 210 &gt; 1 &lt; 211 &gt; 243 &lt; 212 &gt; PRT &lt; 213 &gt; Homo sapiens &lt; 4 Ο Ο &gt; 1

Asp Glu Pro Pro Gin 5 Ser Pro Go Tyr Go Asp 20 Go Leu Lys Phe Glu Gly 35 Ser Ala Leu Sly Asu Trp Asp Ser Go Thr Ser 50 55 Gly Pro Go Thr Gin Glu Phe 65 70 Gly Leu Arg Glu Glu Met Ser 85 Go to GJ.P Pro Tyr Leu Asp ASP 100 Glu Leu Tyr Arg Gin Lys Val 115 Gly Ala Arg Gin Lys Leu BlS 130 13S eiy Glu Glu «et Arg Asp Arg 145 150

Arg Vai Lys Asp Leu Ala Thr 10 15 Gly Arg Asp Tyr Will Be Gin 30 Leu Asn Leu Lys Leu Leu Asp

4S

Ser Lys Leu Arg Glu Gin Laws 6D

Asn Leu Glu Lys Glu Thr Glu 75 80

Leu Glu Glu Vâl Lys Ala Lys 90 95

Lys Lys Trp Gin Glu Glu Het 110

Leu Arg Ala Glu Leu Glu Glu 125

Gin Glu Lys Leu Ser Pro Leu 140

Ala Bis Go Asp Ala Leu Arg 155 160 108 Thr Leu. Ala Pm Tyr Ser Asp eia leu Arg Gin. Arg 165 270 Arg Leo. AIA Leu Lys Giu Asn Gly Gly Am ISO 185 His Ala Lys Ala Thr Glu His Leu Ser Thr Leu 185 200 Pro Ala Leu Glu Asp Leu àtq Gin Gly Lee Leu

Leu Wing 175

Arg Leu Sêr Glu 205Pro will

Wing Glu Tyr ISO

Lys Alâ Lys

Leu Glu Ser Λ * 10 22Õ

Phe Lys Will Be Phe Leu Be Ala Leu Glu Glu Tyr Thr Lys Lys Léu 225 230 235 '240

Asn Thr Gin

&lt; 210 &gt; 2 &lt; 211 &gt; 244 &lt; 212 &gt; PRT &lt; 213 &gt; Homo sapiens &lt; 220 &gt; &lt; 221 &gt; MISC_FEATURE &lt; 222 &gt; (1). (1)

&lt; 223 &gt; Cys at the N-terminus &lt; 220 &gt; &lt; 221 &gt; MISC_FEATURE &lt; 222 &gt; (2) .. (244) &lt; 223 &gt; Amino Acids No. 25 to 267 Human Apo A1 &lt; 400 &gt; 2 109

Cys Ãsp Glv Pr o Pro Gin Ser Prpp Asp Arg Val Lys Asp leu Ala 1 5 10 15 tJftr Val Tyr Aap is Leu Lys Asp Ser Gly Arg Asp Tyr Will Be 20 25 30

Gin Phe Glu 61 and Ser Ala Leu 6th Lys Gin Leu Asa Leu Lys Leu Leu 35 m 45

Asp Asn Trp Asp Will Be Thr Be Thr Phe Be Lys Leu Arg Gla Gin 110 55 60

Leu Gly PIO Vâl Thr Gin GIu Phe Trp Asp Asa Leu Glu Lys Glu Thr 65 70 75 &quot; 80 01¾ Gly Leu Arg Gin Glu Met Ser Lye Asp Leu Glu Glu Val Lys Ala §5m 95

Lys Go Gin Pro Tyr Leu 100

Asp Phe Gin Lys Lys frp Gin Glu Glu 105 1.10

Met Glu Leu Tyr Arg Gin Lys Go Glu. Pr o Leu Arg Ala Glu Leu Gin 115 '120 125

Glu Gly Ala Arg Gin Lys Leu Mis Glu Leu. Sln Glu Lys Leu Ser Pro 130 135 140

Leu Gly Glu Glu Ksfc 145 150

Arg Ala Arg Ala Bis Go Asp Ala Leu 155

Arg Thr Bis Leu Wing Pro Tyr Ser Asp Glu Leu Arg Gin Arg Leu Wing 165 170 175

Ala Arg Leu Glu Ala Leu Lys Glu Asa Gly Gly Ala Arg Leu Ala Glu 185 im

Tyr Bis Ala Lys Ala Thr Glu Bis Leu Ser Thr Leu Set Glu Lys Ala 195 200 205

Lys Pro Ala Leu Glu

Leu Arg Gin Gly Leu Leu Fro Vâl Leu Glu 215 '220

Ser Fhe Lys Will Be Phe Leu Be Lea Leu Glu Glu Tyr fhr Lys Lys 225 230 235 240

Leu Asa Thr Gin 111

&lt; 210 &gt; 3 &lt; 211 &gt; 301 &lt; 212 &gt; PRT &lt; 213 &gt; Homo sapiens &lt; 220 &gt; &lt; 221 &gt; MISC_FEATURE &lt; 222 &gt; (1) .. (58) &lt; 223 &gt; Trimerization module from tetranectin &lt; 220 &gt; &lt; 221 &gt; MISC_FEATURE &lt; 222 &gt; (59). . (301) &lt; 223 &gt; Apo Al mature &lt; 400 &gt; 3 112 112 Thr Gin Lys 10

Ser Pro Gly Thr G.iu Fr Pro 1 S

Pro Lys Lys II® Go Asa 15

Ala Lvs Lys Asp Go Go Asri 20

Thr Lys Het 25

Phe Glu Glu Leu Lys Ser 30

Arg Leu Asp Thr Leu Ala Gin 3 $ 61u Go Ala 40 '

Leu Leu Lys Glu Glu Gin 45

Ala Leu Gin Thr Will Be Leu 50 55

Lys Gly Ser Asp Glu Pro Pro Gin Ser 60

Pro Trp Asp Arg Val Lys Asp Leu Ala Thr Val Tyr Asp Vai Leu 65 70 75 80 Lys Asp Ser Gly 85 Asp Tyr Will Be Glu 90 Phe Glu Gly Ser Ala 95 Leu Gly Lys Gin Leu Agn Leu Lys LSU Leu Asp Asn Trp Asp Ser is Thr 100 105 110 Ser Thr Pha Ser Lys Leu Arg Glu Gin Leu Gly Pro Go Thr Gin Glu 115 120 125 Fhe Trp Asp Asn Leu Glu Lys Glu Thr Glu Gly Leu Arg Glu Glu Met 130 135 140

Ser Lys Asp Leu Glu Glu Go Lys Ala Lys Go Gin Pro Tyr Leu 145 150 155 ISO

Phe Gin Lys Lys Trp Gin Glu Glu Mèt Glu Leu Tyr Arg Gin Lys 165 HO Π 5 goes Glu Pro Leu Arg Ala Glu Leu Gin Glu Gly Ala Arg Gin Lys Leu 113 180 185 190

My Gltt Leu Gin. Gin Lys Leu Ser Pro Leu Gly 31u Glu Met Arg Mp 195 200 3:05

Arg Ala Arg Ala His Vai top Ala Leu Arg fhr Kis Leu Ala Pro Tyr 210 21 $ 220

Ser Asp Glu Leu. Arg Gin Arg Leu Ala Ala Arg Leu Glu Ala Leu Lys 225 230 23 $ 240

Glu Asft Gly Gly Ala Arg L # u Ala Glu Tyr My Ala lys Alâ Thr Glu 24 $ 250 2S $

His Leu Ser Thr Leu Ser Glu Lye Ala Lys Pro Ala Leu Glu Asp Leu 260 26 $ 270

Arg Gin Gly Leu Leu Pro Vsl Leu Glu Ssr Phe Lys Will Be Phe Leu 27 $ 280 28 $

Ssr Ala Leu Glu Glu Tyr Thr Lys Lys Leu Asn Thr Gin 290 295 300

&lt; 210 &gt; 4 &lt; 211 &gt; 258 &lt; 212 &gt; PRT &lt; 213 &gt; Homo sapiens &lt; 220 &gt; &lt; 221 &gt; MISC_FEATURE &lt; 222 &gt; (1) .. (58) &lt; 223 &gt; Trimerization module from tetranectin &lt; 220 &gt; 114 &lt; 221 &gt; MISC_FEATURE &lt; 222 &gt; (59) .. (258) &lt; 223 &gt; Amino acids Apo-68-267 human &lt; 400 &gt; 4

Ser Pro Gly Thr Glu Pro Pro Thr 1 5

Gin Lys Pro Lys Lys Ile 1.0

Lys Lys Asp Go To Asa Thr. 20

Lys Phe Glu Glu Leu 25 30

Will be 15 Lys Ser 115

Arg leu Asp Ihr Leu Me G.ln Glu Vai Ais Lsu Leu Lys Glu Gin &lt; 51n 3S 40 45

Ala Leu Î ± Î ± ββββγβββγβγγβγγγγγγγγγγΠSerÎ SerÎ SerÎ AsÎ AsÎ AsÎysÎysÎysÎysÎysys Le Le Le Le Le Lee Lee Lee Lee Lee Lee l Le Le Lee Lee Lee Lee Lee Lee Lee Lee Lee Lee Lee Lee Lee Lee Lee Lee Lee Lee Lee Lee Lee Lee Lee Lee Lee Lee Lee Lee Lee Lee Lee Lee Lee

Pro Glu Phe Trp Asp Ase Leu 61 «Lys siu Thr Slts s.ly 85 50 35

Leu Arg Gin 61 «Ket Ser Lys Asp Leu 61 or 61« Go »* ys Ma Lys Go 100 105 Á1Q

Gin Pr © Tyr Leu A $ p Asp Phe Gin Lys Lys? Xp Gla Glu Glu .Met Glu Π5 ISO 335 tea Tyr Àrç Gin. Lys Vai G.la Pro Leu Arg Ala Glu Leu Gin Glu Gly 130 135 140

Ala Arg Gin Lys Leu Mie Glu Leu Gin Glu lys Leu Ser Pr © Leu Gly 145 150 155 160

Gla Gla Met Arg Aap Arg Aa Arg Ala Kis Vai Agp Ala Leu Arg T &amp; 365 1701 175

His Les Ala Pr © Tyx Ser Asp Glu Le «Arg Gin Arg Leu Ale Ala Arg ISO 185 190

Leu Gla Ala tu Lys Glu Asa Gly Gly Ala Arg Leu Ala Glu Tyr His 135 200 '205

Ala Ly $ Ala Thr Gla His Ltu Ser Thr Leu Ser 6.1¾ Lys Ala Lys Pro 210 215 220

Ala Leu Glu Asp Leu Arg Gin Gly Leu Leu Pr © Vai Leu Glu Set Phe 225 230 235 240

Lys Will Be Phe Leu Be Ala Leu Glu Glu Tyr Thr Lys Lys Leu Asn 245 250 2 $

Thr Gin 116

&lt; 210 &gt; 5 &lt; 211 &gt; 301 &lt; 212 &gt; PRT &lt; 213 &gt; Homo sapiens &lt; 220 &gt; &lt; 221 &gt; MISC_FEATURE &lt; 222 &gt; (1) .. (58) &lt; 223 &gt; Trimerization module from tetranectin &lt; 220 &gt; &lt; 221 &gt; MUTAGEN &lt; 222 &gt; (9) .. (9) &lt; 220 &gt; &lt; 221 &gt; MUTAGEN &lt; 222 &gt; (15). . (15) &lt; 220 &gt; &lt; 221 &gt; MISC_FEATURE &lt; 222 &gt; (59). . (301) &lt; 223 &gt; Apo-Al &lt; 400 &gt; 5 117

Ser Pro Gly Thr Glu Pro Pro Thr Gin Lys Pro Lys Ala IIe Go Asn 1 5 10 15

Ala Lys Ala Spp Vai Vai Aso Thr Lys ttet Phe Giu Glu Leu Lys Ser 2Q 25 30

Arg Leu Asp Thr Leu Ala Gin Glu Vai Ala Leu Leu Lys Glu Gin Gin 35 40 45

Wing Leu Gin Thr Will Be Leu Lys Gly Ser Asp Glu Fro Pro Gin Ser 50 55 '60

Pro Trp Asp Ax Go Lys Asp Leu Ala Thr Go Tyr Go Asp Go Leu 65 70 75 80

Lys Asp Ser Gly Arg Asp Tyr Will Be Gin Phe Glu Gly Ser Ala Leu 85 â S S5

Gly Lys Gin Leu Asn Leu Lys Leu Leu Asp Asn Trp Asp Ser Vâl Thr 100 105 110 118 be Thr Fbe Ser lys Leu Arg Glu Gin Leu Gly 11.5 120

Pro V Thr € -1 r. Glu 125

Asp Leu (31 u Lys Siu Thr Glu 130 135

Leu Arg Gin Glu Met 140

Ser Lys Asp Leu Giu Gin Go Lys Wing Lys Go Gin Fro Tyr Leu Mp 1.4 S 150 155 160

Asp Phe Gin Lys Lys Trp Gin Glu Glu Giu Leu Tyr 165 170

Gin Lys 175

Go Gin Pto Leu .Arg Wing Glu Leu Gin Glu Gly Wing Arg Gin Lys Leu ISO * 13S 190

Hia Glu Leu Gin Glu Lys Leu Ser Pro Leu Gly Giu Glu Met Arg Asp 195 200 205

Arg Ala Arg Ala Ris Go Asp Ala Leu Arg Thr Bis Leu Ala Pro Tyr 210 215 220

Ser Asp Glu Leu 225

Gin Arg Leu Ala Ala Arg Leu Glu Ala Leu Lys 230 235 240

Glu Asn Gly Gly Ala Arg Leu Ala Glu Tyr Hi Ays Lys Ala Thr Glu 245 &quot; 250 255

His Leu Ser Thr Leu Ser Glu Lys Ala Lys Pro Ais Leu Glu Asp Léu 260 255 270

Arg Gin Gly Leu Leu Pro Vai Leu Glu Be Phe Lys Will Be Fhe Leu 275 280 '285

Ser Wing Leu Glu Glu Tyr Thr Lys Lys Leu Asn Thr Gin 230 295 300 &lt; 210 &gt; 6 119

&lt; 211 &gt; 304 &lt; 212 &gt; PRT &lt; 213 &gt; Homo sapiens &lt; 220 &gt; &lt; 221 &gt; MISC_FEATURE &lt; 222 &gt; (1) .. (58) &lt; 223 &gt; Trimerization module from tetranectin &lt; 220 &gt; &lt; 221 &gt; MISC_FEATURE &lt; 222 &gt; (59). . (61) &lt; 22 3 &gt; Linker &lt; 220 &gt; &lt; 221 &gt; MISC_FEATURE &lt; 222 &gt; (62). . (304) &lt; 223 &gt; Mature Apo-AI &lt; 400 &gt; 6 120 120 s lie Go Asn 15

Ser Pro Gly Thr Glu Pr Pro Thr Gin Lys Pr o Lys l 5 10

Ala Lys Lys Asp Go Go Asn Thr Lys Met Fhe Glu Glu Leu Lys ser 20 25 30

Arg Leu Asp Thr Leu Ala GXn Glu Vai Ala Leu Leu Lys Glu Gin Gin 35 40 45

Ala 'Leu Gin Thr Will Be Leu Lys Gly Be Be Gly His Asp Glu Pru 50 55 60

Prc < Gin Ser Pro Trp Asp Arg Go Lys Asp Leu Ala Thr Go Tyr Go 65 70 75 S0

Leu Lys Asp Ser Gly Arg Asp Tyr Will Be Gin Phe Glu Gly 85 90 95 Ala Leu Gly Lys Gin Leu Asn Leu Lys Leu Leu. Asp Asn Trp hSp 100 105 110

Being Will Be Thr Will Be Lys Leu Arg Glu Gin Leu Gly Pre Go 115 120 125

Thr Gin Glu Phe Trp Asp Asn Leu Glu Lys Glu Thr Glu Gly Leu Arg 130 '135' 140

Gin Glu Met Ser Lys Asp Leu Glu Glu Vai Lys Ala Lys Vai Gin Prc 145 150 155 160 fyr Leu Asp Asp Phe Gin Lys Lys Trp Gin Giu Glu Met Glu Leu Tyr 165 170 175 121

Arg Gin Lys Go Glu Piro Leu Aro Ala Glu Leu Gin Glu Gly Ala Arg im iBB iso

Gin Lys Leu His Glu leu Gin Giu Lys Leu Ser Pro Leu Gly Giu Glu 193 200 '205

Mefc Arg Aβ p Arg Ala Arg Ala Bis Va Asp Ala Leu Arg Thr His Leu 210 '215 220

Ala Pr o fyr Ser Asp Glu Leu Arg Gin Arg Leu Ala Ala Arg Leu Glu 225 230 235 '' 240

Ala Leu Lys Glu Aso Gly Gly Ala Arg Leu Ala Glu Tyr His Ala Lys 245 250 255

Ala Thr Glu Bis Leu Ser Thr Leu Ser Glu Lys Ala Lys Pto Ala Leu 260 265 210

Glu Asp Leu Arg Gin Gly Leu Leu Pr © Vai Leu Glu Ser Phe Lys goes 275 280 285

Ser Phe Leu Ser Ale Leu Glu Glu Tyr Thr Lys Lys Leu Asn Thr Gin 290 295 '&quot; 300

&lt; 210 &gt; 7 &lt; 211 &gt; 304 &lt; 212 &gt; PRT &lt; 213 &gt; Homo sapiens &lt; 220 &gt; &lt; 221 &gt; MISC_FEATURE &lt; 222 &gt; (1) .. (56) &lt; 223 &gt; Trimerization module from tetranectin &lt; 220 &gt; 122 &lt; 221 &gt; MISC_FEATURE &lt; 222 &gt; (57) .. (61) &lt; 223 &gt; Linker based on fibronectin &lt; 220 &gt; &lt; 221 &gt; MISC_FEATURE &lt; 222 &gt; (62). . (304) &lt; 223 &gt; Mature Apo-AI &lt; 400 &gt; 7

Peo Gly Thr Gly Pro Pre Thr Gin Lys? Ro Lys Lys Xle Go Asn 123 15 10 15

Ala Lys hf $ Aap Goes Asn Thr Lys Mêt Phe 61 «Glu Leu Lys S« r 2ϋ 25 30 A: rq Leu Asp Thr Leu ftla sln Gl «vai Ala Leu Leu Lys Glu Gin Gin 35 4D 45 Bad Leu Gla Thr Go Ser Leu Lys Sly Thr Ser Gly Gin Asp Gin Fro 50 55 60

Fro Gin Ser Pro Trp ftsp Arg v.al Lys Asp Leu Ala Thr Vai Tyr Go S5 70 75 80

Aso Vai Leu Lys Asp Ser Gly Arg Asp Tyr Will Be Gin Efte Siu SXy. so 35

Be Wing Leu Gly Lys Gin Leu Asn Lau Lys Leu Leu Asp Asn Trp Asp 100 '105 U0

Being Will Be Thr Will Be Lys Leu Krg Glu Gin Leu Gly Pr © Vai 115 120 125

Thr Gin Glu Phe Trp. Asp Asn Leu Giu lys <51u Thr 61 «Gly Leu Arg 130 135 140

Gin Glu Met Ser Lys Asp Leu Glu Glu. Go Lys Ala Lys Go Gin Fro 145 "1-50 155 160

Tyr Leu Asp Asp Phe Gin Lys Lys Trp Gin Glu Glu Met Glu Leu Tyr 165 170 175

Arg Gin Lys Go Glu Fro Leu Arg Ais Glu Leu Gin Glu Gly Ala Arg ISO 18.5 190

Gin Lys Leu Ris Glu Leu Gin Glu Lys thy Be Pr Leu Gly Glu Glu 193 200 205 het Arg Asp Arg Ala Arg Ala His Vai Asp Ala Leu Arg Thr His Leu 210 215 220

Ala pr © Tyr Ser Asp Glu Leu Arg Gin Arg Leu Ala Ale Arg Leu Glu 225 230 '235 240 124

Ai ® Leu Ly ® Glu Aâri Gly Gly Alâ Alâ Glu Tyr Bis Ala Lyj 24 $ 2SÕ 255

Ala Thr Glu Mis Leu Ser Tiir Leu Ser Glu Lys Ala Lys Fro Ala leu 260 265 270

Glu Mp Leu Arg Gin Gly lati Leu Pr © Vai Leu Glu Ser Phe Lys Go 215 280 285

Ser Phe Leu Ser Ala Leu Glu Glu. Tyr fhr Lys Lys Leu Asn Thr Gin 2.90 295 500

&lt; 210 &gt; 8 &lt; 211 &gt; 304 &lt; 212 &gt; PRT &lt; 213 &gt; Homo sapiens &lt; 220 &gt; &lt; 221 &gt; MISC_FEATURE &lt; 222 &gt; (1) .. (56) &lt; 223 &gt; Trimerization module from tetranectin &lt; 220 &gt; &lt; 221 &gt; MUTAGEN &lt; 222 &gt; (13) .. (13) &lt; 220 &gt; &lt; 221 &gt; MUTAGEN &lt; 222 &gt; (19).

&lt; 220 &gt; &lt; 221 &gt; MISC FEATURE 125 &lt; 222 &gt; (57) .. (61) &lt; 223 &gt; Linker based on fibronectin &lt; 220 &gt; &lt; 221 &gt; MISC_FEATURE &lt; 222 &gt; (62) .. (304) &lt; 223 &gt; Mature Apo-AI &lt; 400 &gt; 8

Ser Pro GXy Thr G1 or Pro Pr © Thr Gin. Lye Prs Lys Ma II © Go Asn 1 5 10 U

Wing Lys Wing A.sp Goes Asrs Thr Lys Het Phe Glu Glu Lsu LyS SâE 20 '25 30 126 htg leu Asp Thr tu Ma Gin Gin will row Leu Leu Lys Glu Gin Gin 35 49 45

Ala Leu Gin Thr Vai Sôr Lea Lys Gly Thr Ser Gly Gin Asp Glu Pro 50 55 '€ 0

Pro Gin Ser Pro Trti Asp fira vai Lys Asp Lea Ala Thr Vai Tyr Go 65 '70 75 80

Asp Go Leu Lys Asp Ser Gly Ar <j Asp Tyr Will Be Gin Mie Glu Gly 95 ^ 95

Be AXa Leu Gly Lys Gin Leu Asn Lea Lys Leu Lea Asp Asn Trp Asp 100 105 110

Being Will Be Thr Will Be Lys Lea Arg Glu Gin Leu Giy Pro Go 115 115 125

Thr Sln Glu Pbe Trp Asp Asn Leu Glu Lys Glu Thr Glu Gly Leu Arg 130 135 140

Gin Glu Mst. Ser Lys Asp Leu Glu Glu Go Lys AXa Lys Go Gin Pro 145 ISO 155 160

Tyr Laa Asp Asp Phe Glu Lys Lys Trp Gin Glu Glu Met Glu Leu Tyr US 1 170 175

Arg Gin Lys Go Glu Pro Leu A * g Ala Glu Leu Gin Glu Gly Ala Arg 190 185 190

Gin Lys Leu His Glu Leu Gin Glu Lys Leu Ser Pro Leu Gly Glu Glu 135 200 '205

Hat Arg Asp Arg Ala Arg Ala His Asp Ala Leu Arg Thr His Leu 210 215 '

Ala Pre Tyr Ser Asp Glu Leu Arg Gin Arg Leu Ala Ala Arg Leu Giu 225 230 235 248

Ala Leu Lys Giu Asn Gly Gly Ala Arg Leu Ala Glu Tyr His Ala Lys 245 250 255

Ala Thr Glu His Leu Ser Thr Leu Ser Giu Lys Ala Lyh Prò Ala Leu 127 260 2 € 5 270

Glu Asp Leu Arg Gin Gly Leu Leu Pro Vla Leu Glu Ser Phe Lye Vai 275 '280 285

Ser Phe Leu Ser Ma Leu Glu Giu Tyr fiir Lye Lys Leu Asn thr Gin 2 »0 295 300

&lt; 210 &gt; 9 &lt; 211 &gt; 306 &lt; 212 &gt; PRT &lt; 213 &gt; Homo sapiens &lt; 220 &gt; &lt; 221 &gt; MISC_FEATURE &lt; 222 &gt; (1) .. (58) &lt; 223 &gt; Trimerization module from tetranectin &lt; 220 &gt; &lt; 221 &gt; MISC_FEATURE &lt; 222 &gt; (59) .. (63) &lt; 223 &gt; Linker &lt; 220 &gt; &lt; 221 &gt; MISC_FEATURE &lt; 222 &gt; (64) .. (306) &lt; 223 &gt; Mature Apo-AI &lt; 400 &gt; 9 128

Ser Fro Gly Tbx Glu Pro Pr © Thr Gin. Lys Pro Lys Lys Ile Val Asn 1 5 1.0 15

Ala Lys Lys Asp Go Go Asn Thr Lys Ifet Phe Glu Glu Leu Lys Ser 20 25 30

Ar§ Leu Asp Thr Leu Ala Gin Glu goes Ala Leu Lau Lys GIu Gin Gin 35 4D 45

Wing Leu Gin Thr Will Be Leu Lys Gly Be Lys Will Sis Het Lys Asp 50 55 '60

Glu Fro Pr © Gin Ser Pr Trp ftsp Arq Vai Lys Asp Leu Ala fhr Go 65 70 15 80

Tyr Go Asp Go Leu Lys Asp Ser Gly Arg Asp Tyr Gonna Be Gin Phe 85 90 §5 129

Gla Gly Set Wing read Gly Lys Gin Leu Ase Leu Lys Leu Leu Asp Ase 100 XOS 110

Trp Asp Ser Thr Ser Thr Ser Thr Lys Leu Arg Gla Gin Leu Gly 115 120 '125

Pr © Go Thr Gin Glu Pise Trp Asp As-n Leu Glu Lys Gla Thr Glu Gly 130 135 140

Leu Arg Gin Glu Met Ser Lys Asp Leu Glu Glu Vai Lys Ale Lys Tal 145 150 155 ISO

Gin Pro Tyr Leu Asp Asp Phe Gin Lys Lyâ Trp Gltt Glu Glu Hat Gla 1SS 130 Π5

Leu Tyr Arg Gin Lys Go Gl.u Prc < Leu Arg Ala Glu Leu Gin Glu Gly 180 1 & 5 190

Ala Arg Gin Lys Leu His Glu Leu Gin Glu Lys Leu Ser Pr © Lsu Gly 195 200 205

Glu Glu Hat Arg Asp Arg Ala Arg Ala His Vai Asp Ala Leu Arg Thr SIS 215 * 220

His Leu Ala Pro Tyr Ser Asp Glu. Leu Arg Gin Arg Leu Ala Ala Arg 225 230 235 240

Leu Glu Ala Leu Lys Glu Asn Gly Gly Ala Arg Leu Ala Glu Tyr His 245 250 225

Ala Lys Al «Thr Glu His Leu Ser Thr Leu Ser Glu Lys Ala Lys Pro 2S0 .265 270

Ala Leu Glu Asp Leu Arg Gin Gly Leu Leu Pro Vla LfcU Glu S r rhe he 275 1 280 285

Lys Vai Ser PAe Leu Ser Aia Léu Glu Glu Tyr Thr Lys Lys Leu Asn 230 295 300

Thr Gin 305 130 &lt; 210 &gt; 10 &lt; 211 &gt; 306 &lt; 212 &gt; PRT &lt; 213 &gt; Homo &lt; 22 &gt; &lt; 221 &gt; MISC_FEATURE &lt; 222 &gt; (1) .. (56) &lt; 223 &gt; Trimerization module from tetranectin &lt; 220 &gt; &lt; 221 &gt; MISC_FEATURE &lt; 222 &gt; (57). . (63) &lt; 223 &gt; Linker based on tetranectin &lt; 220 &gt; &lt; 221 &gt; MISC_FEATURE &lt; 222 &gt; (64) .. (306) &lt; 223 &gt; Apo-Al &lt; 400 &gt; 10 131

Ser Ρε © Gly Thr Glu Pro Pro Thr Gin Lys Pro lys Lys lie Val Asn 1 5 10 15

Allah. Lys Lys Asp Go To Aon Thr Lys Met Phe 61u Glu. Leu Lys Ser 20 25 30

Mg Leu Asp Thr Leu Ala Gin Glu Vai Ala Leu Leu Lys Glu Gin Gin 35 40 45

Leu Gin Thr Will Be Leu Lys Gly Thr Lys Will His Met Lys Asp 50 55 60

Glu Pr Pro Pro Ser Trp Asp Arg Val Lys Asp Leu Ma Thr Val 65 10 75 75 80

Tyr Will Asp Go Leu Lys Asp Ser Gly Arg Asp Tyr Will Be Gin Fhe 85 9θ 95

Glu Gly Ser Ala Leu 1.00

Lys Gin Leu Asn Leu Lys Leu Leu Asp A $ n 105 110? Rp Asp Be Will Thr Be Thr Phe Be Lys Leu Mq 61 «Gin Leu Gly 115 120 125

Pro Go Thr Gin Glu Phe Trp Asp Asn Leu Glu Lys Gin. Thr Glu. Gly 130 135 140 132

Leu Arg Gin Glu Met Ser Lys Asp Leu Glu Gin Go Lys Ala Lys Go 145 145 150 160

Gin Pro Typo leu Mp Asp F.he SI »Lys lys Trp Sl» Glu Glu Het Giu 165 Π0 175 leu Tyi Arg Gin Lys Go Glu Ptú Leu Ara Alâ Gin Leu Gin Giu Gly 180 185 190

Ala Arg Gin Lys Leu His Glu Leu Gin Glu Lys Leu Ser Pro Leu Gly 195 '200' 205

Glu Glu phosphat Arg Asp Arg Ala Arg Ala His Vai Asp Ala Arg Arg 210 215 215

Bis leu Alâ Pro fyr Ser Asp Glu Leu Arg Gin Arg Leu Ala Alâ Arg 225 230 235 240

Leu Glu Ala Leu Lys Glu Asn Gly Gly Ala Arg Lei Ala Glu Tyr His .24 5 &quot; 250 255

Allah. Lys Ala Thr Glu His Leu Ser Thr Leu Ser Glu Lys Ala. Lys Pro 260 2:65 270

Wing read Glu Asp Leu Arg Gin Gly Lau Leti Gold Go Leu Glu Ser Phe 275 .280 285

Lys Will Be Phe Leu Be Ala Leu Glu Glu fyr Thr Lys Lys Leu Asn 290 295 300

Thr Gin 305 &lt; 210 &gt; 11 &lt; 211 &gt; 306

&lt; 212 &gt; PRT &lt; 213 &gt; Homo sapiens &lt; 220 &gt; &lt; 221 &gt; MISC_FEATURE &lt; 222 &gt; (1) .. (56) &lt; 223 &gt; Trimerization module from tetranectin &lt; 220 &gt; &lt; 221 &gt; MUTAGEN &lt; 222 &gt; (13) .. (13) &lt; 220 &gt; &lt; 221 &gt; MUTAGEN &lt; 222 &gt; (19) .. (19) &lt; 220 &gt; &lt; 221 &gt; MISC_FEATURE &lt; 222 &gt; (57). . (63) &lt; 223 &gt; Linker based on tetranectin &lt; 220 &gt; &lt; 221 &gt; MISC_FEATURE &lt; 222 &gt; (64) .. (306) &lt; 223 &gt; Apo-Al &lt; 400 &gt; 11 134

Ser Pro Gly Thr Siu Pro Pro Thr Gin Lys Pro Lys Ala Ile Val Asn 1 5 Ú * 15

Ala Lys Ala Asp Go Go Asn Thr Lys Met Phe GIu Giu Leu Lys Ser 20 25 30

Arg Leu Asp Thr Ltu Ala Gin Gin Goes Ala Leu tu Lys Giu Gin Gin 35 40 45

Wing Leu Gin Thr Will Be Leu Lys Gly Thr Lys Will His Met Lys Asp 50 55 m

Glu Pro Pro Gin Ser Pro Trp Asp Arg Vai. Lys Asp Leu Ala Thr Go m 70 75 80

Tyr Go Asp Go Leu Lys Asp Ser Gly Arg Asp Tyr Will Be Gin Phs 85 θΟ 35

Glu Gly Ser Ala Leu Gly Lys Gin Leu Asn Leu Lys leu Leu Asp Asm 100 '105 110

Trp A $ p Ser V $ 1 Thr Lur Thr Phe Ser Lys LÃ © rg Glu Gin Leu Gly 115 120 125

Pro Go to Thr Gin Glu Phé Trp Asp Asn Leu Glo Lys SIu Thr Glu Gly 130 135 140

Leu Arg Gin GXu Mefc Ser Lys Asp Leu Glu Glu Go Lys Ala Lys Go 135 14.5 ISO 155 160

Gin Pr o Tyr Lau Asp Rsp Phe Gin ly $ Lys Trp Gin Glu Glu Met Glu 165 'Π 0 175

Tyr arg Gin Lys Go Glu Pro Leu Arg Wing Glu Leu Gin Glu Gly 180 185 ~ 190

Allah Ã.rg Gin Lys L &amp; 81 "Glu Leu Glu Glu Lys Leu Ser Pro Leu Giy 195 200 205

Glu Glu Met Arg Asp Arg Ala Arg Ala His Va Asp Ala Leu Arg Thr 210 * 215 220

His Leu Ala Pro Tyr Sar Asp Glu Leu Arg Arg Leu Ala Ala Arg 225 230 230 240

Leu Glu Ale. Lys Glu Asa Gly SXy Ala Arg Leu Ala Glu Tyr Sis 245 250 &quot; 255

Ala Lys Ala Thr Glu His Leu Ser Thr Leu Ser Glu Lys Ala Lys Pro 260 265 '270

Ala Leu Glu Asp Leu Arg Gin 61y Leu Leu Pro · Vai Leu Glu Ser Phe 275 280 285

Lys Will Be Fhe Leu Be Lea Leu Glu Glu Tyr Thr Lys Lys Leu Asn 290 295 .300

Thr Gin. 305

&lt; 210 &gt; 12 &lt; 211 &gt; 51 &lt; 212 &gt; PRT &lt; 213 &gt; Homo sapiens 136 &lt; 3 Ο Ο &gt; &lt; 3 &gt; 2 &gt; Trimerization module &lt; 310 &gt; WO 98/56906 &lt; 311 &gt; 1998-06-11 &lt; 312 &gt; 1998-12-17 &lt; 313 &gt; (1) .. (51) &lt; 400 &gt; 12

61a Pro Thr Gin Lys Pro Lys Lys Ile Asn Ala Lys Lys Asp 1 5 10 IS

Go Go Wing Thr Lys Met Phe Glu Giu Leu Lys Ser Arg Leu Asp Thr 20 25 30

Leu Mâ Gin Gia Go Lea Leu Leu Lys Glu Gin Gin Lea Gin Thr -35 40 45

Go C ys Leu 50 &lt; 210 &gt; 13 &lt; 211 &gt; 58 &lt; 212 &gt; PRT &lt; 213 &gt; Homo sapiens &lt; 220 &gt; &lt; 221 &gt; MISC_FEATURE &lt; 222 &gt; <22> <223> <223> <223> <223> <223> <223> <223> <223> <223> </ (58) <22 3> Linker &lt; 400 &gt; 13 &lt; 222 &gt; MISC_FEATURE &lt;

Ser Pro Sly Thr çiu Pro Pro Thr Gin Lys Pro Lys Lys lie Go Asr &gt; 15 10 15

Ala Lys Lys Asp Go Vai Asn Thr Lys Hefc Phe Glu 61 α Leu Lys Ser 20 25 30 Âμg Asp Thr Leu Ala Gin Glu Vai Ala Leu Leu Lys 61 or Glrs Gin 40 35

Ala Leu Gin Thr Will Be Leu Lys Gly Ser 50 55

&lt; 210 &gt; 14 &lt; 211 &gt; 329 &lt; 212 &gt; PRT &lt; 213 &gt; Homo sapiens &lt; 220 &gt; &lt; 221 &gt; MISC_FEATURE &lt; 222 &gt; (1) .. (86) &lt; 223 &gt; Hp (alpha) residues 138 &lt; 22 &gt; &lt; 221 &gt; MISC_FEATURE &lt; 222 &gt; (87) .. (329) &lt; 223 &gt; Αρο Α-Ι &lt; 400 &gt; 14 139

Gly Go Asp Ser Gly Asn Asp Go Thr Asp ile Ala Asp Aβ Gly Cys 1 5 10 15

Pro Lys Pr o Pro 61 u Ile Ala uis Gly Tyr goes Gin His Ser Vai Arg 20 25 30

Tyr Tour Cys Lys Asn Tyr Tyr Lys Leu Arg Thr 61a Sly Asp Gly VAX 35 40 '4 $'

Tyr Thr Leu Asn Asa 61 «Lys Gin Trp Ile Asn Lys .Ala Vai Gly Asp 50 55 '60

Lys Leu Pro 61u Cys 61a Ala Vai Ala Gly Lys Pro Lys Asn Pro Ala 65 70 75 BQ

Asn Pro Vai Gin As: Asp Glu Pro Pro 61a Ser Pro Trp Asp Aro 85 90 SS Vâl Lys Asp Leu Ala Thr 100

Tyr Vâl Asp VÎμ Le &amp; Ly.s Asp Ser Gly 105 110

Ser Ma Lou Sly Lys 61 »Leu 125

Arg Asp Tyr Go to * Gin Phe 61 or 63 115 120

Asn Leu Lys Leu Leu Asp Asn Trp Asp Ser Vl Thr: Ser Thr Phi Ser 130 135 148

Lys Leu Arg Glu Gin Leu Gly Pro Go Thr Gin Glu Phe Trp Asp Asn 145 150 155 160

Leu Gl «Lys Glu Thr Gly Gly Leu Arg Gin Glu Met Ser Lys Asp Leu 140 165 110 175

Glu Slu Vai Lys Ala Lys Sai Pro Tyr Leu Asp Asp Phe Sln Lys 180 185 190 lys Trp Gin Glu Glu Het Glu Leu Tyr Arg Gin Lys Go Glu Pro Leu 195 200 205

Arg Ala Glu Leu Gin Glu Gly Ala Arg Gin Lys Leu His Glu Leu Gin 210 215 '220

Glu. Lys Leu Ser Pro Leu Gly Glu Glu Het. Arg Rsp Arg Ala Arg Ala 225 23Θ 235 240

Mia Vai Asp Ala Leu Arg Thr His Leu Ala Fro Tyr Ser Asp Glu Leu 245 250 255

Arg Gin Arg Leu. Ala Ala Arg Leu Glu Ala Leu Lys Glu Asn Gly Gly 260 265 '270

Ala Arg Leu Ala Glu Tyr His Ala Lys Ala Thr Glu His Leu Ser Thr 275 280 2S5

Leu Ser Glu Lys Ala Lys Fro Ala Leu Glu Asp Leu Arg Gin Gly Leu 290 295 300

Leu Fro Vai Leu Glu Be Phe Lys Be Phe Leu Be Wing Lau Glu 305 31.0 315 320

Glu Tyr Thr Lys Lys Leu Asn Thr Girs 325

&lt; 210 &gt; 15 &lt; 211 &gt; 267 &lt; 212 &gt; PRT &lt; 213 &gt; Homo sapiens &lt; 400 &gt; 15 ith Lys Ala Ala. Vai tmi Thr Leu Ala Go Leu Phe Leu Thr 5 10

Gin Alâ Aeg KiS FM Trp Gin Gin Asp Glu Pr O Fro Gin Ser 61 and Ser is Fro Trp 20 Z 3 142

Asp Arg Go Lys Asp Leu Ma Thr Go Tyr Go Asp Go Leu Lys Asp 35 40 J 41

Ser Gly Acg Asp Tyr Will Be Gin Glu Gly Be Ala Lau Gly Lys 50 55 60

Gin Leu Asn Leu Lvs Leu Leu Asp fts «Trp Asp Ser Thu Ser Thr 65 '70 &quot; 75 80

Fhe Sar Lys Leuven Glu Glu Leu Gly Pt'0 Go Thr GIb 61 «Ph® Trp 85 50 95

Asp Asn Leu 61u Lys 61u Thr Glu Gly Lau Arç Gin 61a Met Ss.r Lys 100 105 Π 0

Asp Le »Glu Glu Go Lys Ala Lys Go Pro Gin Leu Asp Asp Phe 115 120 125

Gin Lys Lys Trp Gin Gin Glu Met 61 «Leu Tyr Arg Gin Lys goes 61to 130 13S 140

Pro Leu Arq Ala Glu Leu qIr Glu 61v Ala Arg Gin Lys Leu Hls 61u 145 '150 &quot; 155 160

Leu Gin Glu Lys Leu Ser Pro Leu Gly Glu Glu Met Arg Asp Arg Ala 165-170 175

Arg Ala Hi Vai Asp Aia Leu Arg Thr His Leu Ala Pro Tyr Ser Asp 180 185 190

Glu Leu Arg Gin Arg Leu Ala Ala Arg Leu Glu Ala Leu Lys Glu Asr 195 200 SOS

Gly Gly Ala Arg Leu Ala Glu Tyr His Ala Lys Ala Thr Glu His Leu 210 215 &quot; 220 S®r Thr Lau ™ 61 "Lys Ala Lys Fro Ala Leu Giu Asp Leu Arg Gin 225 230 235 249

Gly Leu Leu Pro Go Leu Glu Be Ph «Lys Will Be Fhe Leu Be Ala 245 250 255 143

Leu Glu Glu Tyr Thr Lys Lys Leu As.n Thr Gin 260 ^ 265

&lt; 210 &gt; 16 &lt; 211 &gt; 267 &lt; 212 &gt; PRT &lt; 213 &gt; Homo sapiens &lt; 4 0 0 &gt; 16 144 fêefc Lys Ma Ma VaX Leu Thr Leu Ala Go Leu Phe Leu Thr Gly Ser 1 5 10 15

Gin Wing Mg Mia · Phe Trp Gin Gin Asp Glu EJxo Pro Gin Ser Pro Trp 20 25 30

Arg Go Lys Asp Leu Ala Thr Go Tyr Go Asp Go Leu Lys Asp 35 40 40

Ser Gly Arg Asp. Tyr Will Be Gin Phe Glu. Gly Ser Ala Leu. Gly Lys 50 55 60

Gin Leu Asn Leu Lys Leu Leu Aap As »Trp Asp Ser Vai Thr Ser Thr 65 10 75 80

Phe Ser Lys Leu Arg Glu Gin Leu Gly Pro Go Thr Gin Glu Phe Trp 85 ~ * 90 95

Asp Asn Leu Glu Lys Glu Thr Glu Gly Leu Arg Gin Glu Met Ser Lys 100 105 110

Asp Leu Glu Glu Vai Lys Ala Lys VaX Gin Pro Tyr Leu Asp Asp Phe 115 120 125

Gin Lys Lys Trp Gin Glu Glu Met Glu Leu Tyr Arg Gin Lys Go Glu 130 135 140

Pro Leu Arg Ala Glu Leu Gin Glu Gly Ala Arg Gin Lys Leu Bis Glu 145 150 155 160

Leu Glrt Glu Lys Leu Ser Pro Leu Gly Glu Glu Met Arg Asp Arg Ala 165 170 175

Arg Ala His Go Asp Ala Leu Arg fhr .81 * Leu Ala Pro Tyr Ser Asp ISO 185 ISO 145

Slu Leu Arg Qln htg Leu Ala Ala Arg Leu Glu Ala Leu Lys Glu As »195 '200 205

Gly Siy Ala Arg Leu Ala Glu Tyr His Ala Lys Ala Thr Glu His Leu Ϊ20 210? Leu Arg δίπ 240

Ser Thr Leu Sar Glu Lys Ala Lys Pro Ala Leu Glu 225 230 235

Gly Leu Leu Prô Vàl Leu Glu Ser Pfoe Lys Will Be Phe Leu Ser Ala 245 250 255

Leu Glu Glu Tyr Thr Lys Lys Leu Aso Thr Sln 260 265

&lt; 210 &gt; 17 &lt; 211 &gt; 267 &lt; 212 &gt; PRT &lt; 213 &gt; Macaca fascicularis &lt; 400 &gt; 17 146 Lys Ala Thr Vai Leu Thr Leu Ala Vai Leu Phe Leu The Giy Ser 15 10 15

Sln Ala Arg His Phe Tzp Gin Gin & Glu Pro Pro Gin Thr Pro Ttp 20 25 30 Aãp &amp; Uai Lys Asp Leu Vai Thr Vai Tyr Go Glu Ala Leu Lys Asp 35 40 45

Ser Siy Lys Asp Tyr Will Be Gin Phe Glu Siy Ser Ala Leu Giy Lys 50 55 50

Siri Leu Mn Leu Lys Leu Asp Asn Trp Asp Ser Va 1 Thr Ser Thr 65 70 75 80

It will be Lys Leu ftrg Glu Gin Mu Giy Pro Go Thr G.ln Glu Phe Trp 85 SO 95

Asp Asn Leu Glu Lys Glu Thr Glu Giy Leu Arg Gin Glu Met Ser Lys ioo 105 no 147

Asp Leu Glu GIU Go Lys Ma Lys Go Gin Pro Tyr Leu Aap Asp Pha 115 120 125

Gin Lys lys? Rp 61 «Glu Glu Htet Glu Fluro Tyr Mq Gin Lys Vai Glu 130 135 140

Pro Léu Arg Ala Glu Leu Hís Glu Gly Thr Arg Gin Lys Lêu Pis Glu 145 150 155 160

Leu. Bis Glu Lys Leu Ser Pro Leu Gly Glu Glu V Arg Arg Asp Arg Ala 165 170 175

Arg Ala Ma Va.t Asp Ala Leu Arg Thr Sis Leu Ala Pro Tyr Sar Asp 180 185 19α

Glu Leu Arg Gin Arg Lau Ala Ala Arg Leu Glu Ala Leu Lys Glu Asn 195 200 205

Gly Gly Ala Arg Leu Ala 61U Tyr Sis Ala Lys Ala Ser Glu Hys Leu 210 215 220

Ser Thr Leu Ser Glu Lys Ala Lys Pro Ala Leu Glu Asp Leu Arg Gin 225 230 235 240

Gly Leu Leu Pro Go Leu Glu Be Phe Lys Will Be Pha Leu Be Wing 245 250 255

Leu Glu Glu Tyr Thr Lys Lys Leu Ser Thr Gin 260 2:65 &lt; 210 &gt; 18 &lt; 211 &gt; 265 &lt; 212 &gt; PRT &lt; 213 &gt; Bos taurus &lt; 400 &gt; 18 148 mt Lys Ala Go Vai Leu Thr Lee Ala Go Leu P'he Leu Thr Gly Ser 1 5 10 15 61η Ala Arg Hs Phs Trp Gin Gin Asp Asp Pro Gin Ser Ser Trp Asp 20 25 30 Arg Go Lys Asp Pha Ala Thr Val Tyr val Glu Ala 11 «Lys Asp Ser 35 40 45 149

Gly Arg Aep Tyr Go Ala Gin Phe Glu Ala Ser Ala Leu Gly Lys Gin 50 '55 60 Leu Açu Leu Lys Leu Leu Asp pl Trp Asp Thr Leu Ala Ser Thr Leu 65 70 75 80 Ser Lys Go Arg Arg Glu Gin Leu Gly Pro Go Thr Gin Glu Phe Trp Asp 85 90 95 A $ n Leu Glu Lys Glu Thr AM Ser Leu Arg Gin Glu M * t His Lys Asp 100 105 Π0 Leu Glu Glu Go Lys Gin Lys Go Gin Pro Tyr Leu ASp Glu S? Be Glh Π 5 120 125 Lys rys Trp; i .1 ϊϊ Glu Glu Vai Glu lie Tyr Arg Gin Lys Go Ala Pro 130 135 140 Leu Gly Glu Glu Phe Arg Glu Gly Ala Arg Gin Lys Go Gin Glu Leu 145 150 155 160 Gin Asp Lyg Leu Ser Pro Leu Ala Gin Glu Leu Arg Asp Arg Ala Arg 165 170 175 Αίθ Bis Glu Thr Leu Amin Gin Gin Leu Ala Pre Tyr Ser Asp Asp 180 185 100 Leu Arg Gin Arg jj &gt; SU Thr Ala Arg Leu Glu Ala Leu Lys Glu Gly Gly 195 200 205 Gly Ser Leu Ala Glu Tyr His Ala Lys Ala. Ser Glu Gin Leu Lys Ala 210 9 C. 220 Leu Gly Glu Lys Ala Lys Pro Leu Glu Asp Leu Arg Gin Gly Leu 225 230 235 240 Leu Pro Leu Leu Glu Leu Leu Leu Leu Leu Leu Ala Ala Ile Asp 245 250 255 Glu Ala Ser Lys Lys Leu Asn Ala Gin 2S0 265 150 < 210 &gt; 19 &lt; 211 &gt; 265 < 212 &gt; PRT &lt; 213 &gt; Sus scrofa &lt; 4 0 0 &gt; 19 151

Met Lys Ala Go Go Leu Thr Leu Ala Go Lee Phe Leu Thr Giy Ser 15 10 15 51a Ala Arg Bis Phe Trp Gin Gin Asp Asp Pr G Gin Pr Pr Trp Asp 20 2 $ 30

Arg Vai Lys Asp Ph.e Ala Thr Vai Tyr Go Asp Ala Ile Lys Asp Ser 35 40 '45

Giy Arg Asp Tyr Go Wing Gin Pha Glu Wing Be Wing Leu Giy Lys Bis 50 55 00

Leu Asn Leu Lys Leu Leu Asp Asn Trp Asp Ser Leu Giy Ser Thr Phe 65 '70 75 80

Thr Lys Go Arg Glu Gin Leu Giy Pro Go Thr Gin Glu Phe Trp Asp 85 ': 95

Asn Leu Glu Lys Glu Thr Glu Ala Leu Arg Gin Glu Hat Ser Lys Asp 108 10 $ 110

Leu Glu Glu Go Lys Lys Lys Go Gin Pro Tyr Leu Asp Asp Phe Gin 115 120 125

Asn Lys Trp Gin Glu Glu Hat Glu Thr Tyr Arg Gin Lys Met Ala Pro 130 135 140

Leu Giy Ala Glu Phe Arg Glu Giy Ala Arg Gin Lys Go Gin Glu Leu 145 150 155 160

Glu Glu Lys Leu Ser Pro Leu Ala Glu Glu Leu Arg Asp Arg Leu Arg 165 Π0 175

Ala His Goa Glu Ala Leu Arg Gin Goes Ala Pro Tyr Sar Asp Asp ISO '18 $ 190

Leu Arg Gin Arg Met Ala Ala Arg Phe Glu Ala Leu Lys Glu Giy Giy 152 195 200 205

Gly Ser Leu Ma Glu fyr 2 r u

Gin Ala lys Ma Gin Glu Gin leu Lys Wing 215 220

Leu Sly Glu Lys Wing Lys Aro Aia Leu Gin Asp Leu Arq Gin Sly Leu 225 230 235 240

Leu Pr © Vai Leu Glu Asn Leu Lys Vai Ser lie Leu Ala Ala Ile 245 250 255

Glu Ala Ser Lys Lys Leu Assa Ala Gin 250 '265

&lt; 210 &gt; 20 &lt; 211 &gt; 266 &lt; 212 &gt; PRT &lt; 213 &gt; Canis familiaris &lt; 400 &gt; 20 153 Leu Lys Ala Leu Leu Thr Leu Ala Leu Leu Fhe Leu Thr 61 γ Sei 1 5 10 15

Gin Ala Ara 111 s: 20 s Trp Gin Gin Asp <Siu Fro Gin Ser Pro Trp Asp 25 '30

Arg Vai Lys Asp Leu Ala Thr Vai Tyr Go Asp Ala Go Lys Asp Ser 35 40 45 61 and Arg Asp Tyr Vâi Ala Gin Phe Giu Ala Be Ala Leu Gly Lys Gin 50 55 60

Leu Asn Leu Lys Leu Leu Asp Asn Trp Asp Ser Leu Ser Ser Thr Go 65 70 '75 30

Thr Lys Leu Arg Glu Gin Ile Giy Pro Go Thx Gin Glu Phe Trp Asp 85 90 9S Âsn Leu Glu Lys Glu Thr Glu Vai Leu Arg Gin Glu Met Ser Lys Asp 100 105 J 110

Leu Glu Glu Go Lys Gin Lys Go Gin Pm Tyr Leu Asp Asp Phe Gin 115 120 125 154

Lys Lys Trp Gin Glu Glu Vai Glu Leu Tyr Arg Gin Lys goes Ala Pro 130 135 '140

Mu Gly S Glu Leu Arg Glu Gly Ala Arg Gin ty Mu Mu Glu Lôu 145 ISO '155 &quot; 150

Gin Glu Lys Leu Ser Pro Leu Ala Glu Glu Leu Atg Mp Arg Ala Arg 165 170 175

Thr His Vai Asp Ala Leu Arg Ala Gin Leu Ala Pro Tyr Ser Asp A s $ 180 185 190

Leu Arg Glu Arg Leu Ala Ala. Arg Leu Glu Ala Leu Lys Glu Gly &lt; 31 & 195 200 205

Gly Ala Ser Leu Ala Glu Tyr Mie Ala Arg Ala Ser Glu Gin Léu Set 210 215 220

Ala Leu Gly Glè Lys Ma Arg Pro Ala Leu Glu Asp Leu Arg Gin Gly 225 230 235 240

Leu Leu Gold Go Leu Glu Be Phe Lys Will Be Mu Leu Ala Ala lie 245 250 255

Asp Glu. Ala Thr Lys Lyg Leu Asn Ala Glu 2S0 265

&lt; 210 &gt; 21 &lt; 211 &gt; 206 &lt; 212 &gt; PRT &lt; 213 &gt; Oryctolagus cuniculus &lt; 400 &gt; 21 155

Met Lys Ala Vai Leu Thr Leu .Al-â Vai Léu Phe Leu Thr Gly Ser 1 5 10 15 Gin Ala Arg His Phe Trp Gin Arg Asp Glu Fro Arg Ser Ser Trp Asp 20 2S 30 Lys Ile lys Asp Phe Ala Thr Go Tyr Go Asp Thr Go Lys Âμs Ser 35 40 45 Gly Arg Glu Tyr Go to Ala Gin Phe CU: Ala Ser Ai. Ph® Gly Lys Gin 156 50 55 60 read Aan leu Lys leu leu Asp Asn Ίίζφ hsp Ser Leu Ser Ser Thr Go 65 70 75 60

Ser Lys Leu Gin Glu Sln Leu Gly Pro V «1 Thr Gin Gltf Phe Trp Aap

85 90 9S

Asn leu Gin Lys Glu Thr Glu Gly Leu htq Glu Glu Met Asn Lys t uos

Leu Gin Glu Go Arg Arg Lys Go Pro Gin Tyr Leu Asp Glu Phe Gin 115 120 125

Lys Lys Trp Gin Glu Glu Vai Glu Arg Tyr Arg Glu Lys Go Glu Ho 130 135 140

Leu Gly Ala Glu Leu Ara Glu Ser Ala Arg Gin Lys Leu TJxr Glu Leu 145 15Ò 155 160

Glu Glu Lys Leu Sei Fr Leu Ala Glu Glu Leu Arg Asp Ser Ala Arg 165 170 1 1 $

Thr His Go Gly Leu Leu Pro Go Leu Glu Be Phe Lys Will Be Go

180 185 ISO

Gin Asn Vai Leu Asp Glu Ala Thr Lys Lys Leu Mti Tftr Gin 195 200 205

&lt; 210 &gt; 22 &lt; 211 &gt; 265 &lt; 212 &gt; PRT &lt; 213 &gt; Tupaia glis belangeri &lt; 400 &gt; 22 157

Met Lys  ± 3 Go Go Lea ThÃ, ° 1 5 Glrs Ala Arg His Phe Trp Gin Gin 2G Arg Go Arg Agp Leu Ala Asn Go 35 '40

Ala Vai Leu Pfee Leu Thr Gly Ser 10 15 Asp Siu Pr o Gin Ser Ser Trp Asp 25 30 Tyr Go Asp Ala Go Lys 61.ii Ser 15 158

Gly Arg Gia Tyr Go Se * <31 «Leu Lu Ala Se * Ala te» Gly Lys Gin 50 55 60

Leu &amp; Leu Lys Leu Vai &amp; Asp Trp Asp Thr Gly Ser Thr Phe 65 70 75 80

Gin Lys Go H: ls Glu Kis Leu Gly Pr o Go Ala Gin Glu Phe Trp Gin 85 80 SS

Lys Laws Gia Lys Glu Thr Gl »Glu Leu Arg Arg Glu Ile Asm Lys Asp ioo 105 no

Leu Glu Asp Go Arg Gin Lys Thr Gla Pro Fhe Lea Asp Giu Ile Gin 115 &quot; 120 125

Lys Lys Trp Glu Glu. Asp Leu Glu Arg Tyr Arg Gin Lys Go Glu Pro 130 135-140

Leu $ Ala Gin Leu Arg Giu Gly Ala Arg Gin Lys Leu Met Glu Leu 145 15 " 155 160

Gin Glu. Gin Go Thr Pro Leu Gly Glu Asp Leu Arg Asp Ser Vai Arg 165 in &quot; ns

Wing Tyr Wing Asp Thr Leu Arg Thr Gin Leu Wing Pro Tyr Ser Glu Gin 180 1SS 190

Het Arg Lys Thr Leu Gly Ala Arg Leu Glu Ala lie Lys Glu Gly Qly 1.95 200 205

Be Ala Ser Leu Ala Glu Tyr His Ai Lys Ale Be Glu Gin Leu. Ser 210 215 220

Ala Leu Sly Glu Lys Ala Lys Pro Vai Leu Glu .Asp Lie Kis <31 n Gly 225 * 230 "235 240

Read Met P * Het Trp Glu Ser Phe Lys Thr Gly is Leu. Asn Vai lie 245 250 255

Asp Glu Ala Ala Lys Lys Leu Thr Ala 260 265 159

&lt; 210 &gt; 23 &lt; 211 &gt; 264 &lt; 212 &gt; PRT &lt; 213 &gt; Mus musculus &lt; 4 Ο Ο &gt; 23 160

Met Lys Ala Vai Vai Leu Ala Vai Ala Leu Vai Pha Leu Thr Gly Ser 1 5 10 15 Gin Ala Trp Hia Go frp Gin Gin Asp · Glu f Gin Gin Trp Asp 20 25 30 Lys Go Lys Asp Phe Ala Asn Vâl Tyr Go A§p Ala Go Lys Asp be 3.5 40 45 Gly Arg Asp Tyr Will Be Gin Pbe Giu Be Ser Be Leu Gly Without Gin 50 55 60 Leu Asr. Leu Asm Leu Leu Glu Asn Trp Asp Thr Leu Gly Ser Thr Go 65 70 75 80 Ser Gin Lee Gin Glu Arg Leu. Gly Pra Leu Thr Arg Asp Phe Trp Asp 85 90 95 Asn Leu Glu Lys Glu Thr Asp Trp Go Arg Arg Glu Wefc Asm Lys Asp 100 105 110 Leu Glu Gla Go Lys Gin Lys Go Gin Pro Tyr Leu Mp Glu Phe Gin 115 120 125 Lys Lys Trp Lys Glu Asp Go Glu Leu Tyr Arg Gin Lys Go Ala Pra 130 135 140 Leu Gly Ala Glu Leu Gin Glu Ser Ala Arg GXi Lys Leu Gin Glu Leu 145 150 155 16S Gin Gly Arg Leu Ser Pro Vâl Ala Glu Glu Phà © Arg ASp Arg ArgSet Arg 16 $ 170 175 Thr Bis Vs Asp Ser Leu Arg Thr Gin Leu Ala Pro Hls Ser Glu GlR 189 185 190 M «t Arg Giu Ser Leu Ala Gin Arg Leu Ala Glu Leu Lys Ser Asn Pm 195 200 205 161 Thr Leu Asn Glu 210

Tyt Hás Thr ftrg Ala Lys fhr His Leu Lys 21S 230

Thr Leu

Giy Glu Lys Klã Arg Prα Ala Leu Glu Asp Leu Arg filters Ser Leu Mefc 225 '230 235' 240

Leu Glu Tht 245

Leu Lys Thr Lys Ala Gin Ser Vai lie Ace © Lvs 250 255

Ala Ser Glu

thr Leu thr Ala Glí 2 € Q

&lt; 210 &gt; 24 &lt; 211 &gt; 259 &lt; 212 &gt; PRT &lt; 213 &gt; Rattus norvegicus &lt; 400 &gt; 24 162 æl Lys Ai Ala will read Ala Vai Ala leu Vai leu Thr Gly Cys 1 5 10 15 Gin AXâ Trp Glu Phe Trp Gin Gin ASp Glu Pro Gin Ser Gin Trp Aso 20 25 30 Arg Vai lys Asp Phe Ais Tíir Vai Tyr Go Asp Ala Go Lys Asp Ser 35 40 45 siy Arg Asp Tyr Will Be Gln Phe Glu Ser Be Thr Leu Gly Lys Gin 50 55 60 leu Asn Leu Asn leu Leu Ast> Asn Trp Asp Thr Lsu Gly Ser Thr Val 65 70 75 80 Gly Arg leu Gin Glu Gin leu Giy Pr o Go Thr Gin Glu Phe Trp Ala 85 80 05 Asn leu Gin Lys Glu Thr hBP Trp leu Arg Asn. Glu mt Asn Lys ASp 100 105 Π0 Leu GI u ASft Vâl Lys Gin lys Mst Gin Pro Mis Leu Asp Glu Phe Gin 115 120 125 163

Slu Lys Trp Asn Glu 01u Go Glu Ala Tyr Arg Gin Lys Leu Glu Pro 130 135 140

Leu Gly Thr Glu Leu Kis Lys Asn Ala Lys Glu Met Gin ftrg Hís Lêu 145 ISO 155 ISO

Lys Goes Ala Glu Glu Pha Ara Asp Arg Hat Arg Go hm Ala Asp 165 1? 0 175

Ala Leu Arg Ala Lys Phe Gly Leu Tyr Ser Asp Gin Ket Arg Glu Asn 10 105 105

Leu Ala Gin Arg Leu Thr Glu Ile Arg Asn His Pr ohr Leu IX and Glu 195 200 205

Xyr Bis Thr Lys Ala Gly Asp Hys Leu Arg Thr Leu Gly Glu Lys Ala 210 215 220

Lys Pro Ala Leu Asp Asp Leu Gly Gin Gly Leu Met, Pro Vai Leu Glu 225 230 235 240

Ala Trp Lys Ays Lys xle Wet Ser Ket ile Asp Glu Ala Lys Lys Lys 245 250 '255

Leu Wing Wing

&lt; 210 &gt; 25 &lt; 211 &gt; 241 &lt; 212 &gt; PRT &lt; 213 &gt; Erinaceus europaeus &lt; 400 &gt; 25

164 Asp Gin 1 Tyr goes Aep Thr Trp Asp 5Q

Ays Lys Ser Xyr Trp Asp Glft lie Lys Asp 5 10

Met Leu Thr Go 15

Mp Thr Ma Lys à &amp; p Be Giy Lys Asp Tyr20 as

Le · :; Thr Ser Leu 30 Sèr Ala Leu Gly Gin GIb Leu Asa Lys Lys 35 40

Thr Will Be Ser Ala Leu Leu Lys Ala Arcj 55 '60'

Leu Wing Asp Asn 45

Giu Gin Het Lys 165 Pro IIe Wing Wet Glu Phe Ί 65 70 F hsn Leu Glu Lys As 75

Thr Glu Gly 80

Leu Arg Gin Thr Will Be Lys Asp Leu Glu Leu Will Lys Glu Lys Go 85 90 95

Gin Pro Tyt Leu Asp Ser Phe Gin lys Lys Go Glu Glu GM Leu Glu 100 105 110

Leu Tyr Arg Gin Lys Go Ala Pro Leu Ser Ala Glu Trp Arg Glu Gin 115 120 125

Alâ firg Gin Lys Wing Gin GXu Leu Gin Gin Lys Wing Gly Glu Leu Gly 130 135 140

Gin Gin Ris Arg Asp âg Arg Arg Thr His Go Asp Ala Leu Arg Thr 145 150 155 160

Asp Leu A]

Fr or Tyr Gly Glu Glu Ala Arg Lys Leu Leu Leu Gin Arg 165 170 '175

Leu Gin ÃSp Xle Lys Wing Lys Ser Gly Asp Leu Wing Glu Tyr Gin Thi ISO 185 190

Lys Leu Ser Glu His Leu Lys Ser Fhe Gly Glu Lys Ala Gin Pro Thi 195 200 2G5

Leu Gin} 210

Leu Arg Mis Gly Leu Glu Pro Leu Trp Glu Gly Ile 215 220

Ais Gly Ala Met Ser Met Leu Glu Glu Leu Gly Lys Lys Leu Asn Ser 225 230 235 240

Gin &lt; 210 &gt; 26 &lt; 211 &gt; 264 166

&lt; 212 &gt; PRT &lt; 213 &gt; Gallus gallus &lt; 400 &gt; 26 167

Ksfc Arg Gly Go your Vsl Thr Leu Ala Go Leu Phe Leu Thr Gly Thr 1 5 10 15

Gin Ala Arg Ser Phe Trp Gin Ms A * p Glu Fro Yes Thr Pr? Leu Asp 20 25 30

Arg lie Mq Asp Met Yal Asp Go Tyr Leu Q1 e 'Thr Go Lys Ala Ser 35 40 45

Giy Lys Asp Ma lie Ala Sln Mie Glu Ser Ser Ala Go Gly Lys Oln SO '55 60

Leu Asp Leu Lys Leu Ais Asp Asn Leu Asp Thr Leu Ser Ala Wing Wing 65 70 75 80

Ala Lys Leu Arg Glu Asp Met Ala Pr © Tyr Tyr Lys Oiu Val Arg Glu 85 50 §5

Met frp Leu Lys Asp Thr Glu Ala Leu Arg Ala Glu Leu Thr Lys Asp 100 105 110

Leo Glu Glu Go Lys Glu Lys lie Arg Pro Phe Leu Asp Gin Phe Sei 115 I20 125

Ale Lys Trp Thr Glu Glu Leu Glu Gin Tyr Arg Gin Arg leu Thr Pro 130 '135 140

Go Wing Gin Glu Leu Lys Glu Leu Thr Lys Gin Lys Go Glu Leu Met 143 ISO 155 160

Gin Ala Lys L &amp; Thr Pr? Vai Ala Glu Glu Ala Arg Asp Arg Leu Arg 185 17 &quot; 175

Gly Kis Go Glu Glu Leu Arg Lys Asn Leu Ala Tro Tyr Ser Asp Glu 190 1SS '190

Leu Arg Gin Lys Leu Ser Gin Lys Leu Glu Glu Ile Arg Glu Lys Gly 195 200 205

Ha pio Gin Ala Set 'Glu Tyr Glu Ala Lys Vai Het Glu Gin Leu Ser 210 215 220

Asa Leu Arg Glu Lys Ket Thr Pr o Leu Vsl Gin Glu Phg Arg Glu Arg 168 225 230 235 240

Leu Thr Pro Ty.f Ais Glu Asn Leu Ly.s Asn Arg Leu Ϊ1 Ser Ser Leu 245 &quot; 250 255

Aso Glu Leu Glrs. Lys Ser Vai Ala 260

&lt; 210 &gt; 27 &lt; 211 &gt; 264 &lt; 212 &gt; PRT &lt; 213 &gt; Coturnix japonic &lt; 400 &gt; 27 169

Hee Arg 61 and Vai Leu Vai Thr Leu Ala Vai Leu Phe Leu Thr Gly Thr 1 5 10 15 Gin Ala Arg Ser Phe Trp Glu Ris Asp Asp Pro Gin Thr Pro Leu Asp 20 25 30 Arg 11 Âμg ASp Het Leu Asp Vai Tyr Leu Glu Thr goes Lys Ala Ser 35 40 45 Sly Lys Asp Ala 11 «Ser Gin Phe Glu Ser Ser Ala Go Gly Lys Gin S 55 55 Leu Asp Leu Lys Leu Ala Asp Asn Leu Asp Thr Leu Ser Ala Ma Ala 65 70 n 80 Ais Lys Leu Arg eiu Asp Het Thr Pro Tyr Tyr Arg Glu Vai Arg Glu ss 50 95 Mefc Txp Leu *** Thr Glu Mâ Leu Arg Ala Glu Leu Thr Lys Asp 100 105 110 Leu Glu Glu Vai Lys Glu Lys lie Arg Pro Phe Leu Asp Gin The Ser 115 120 125 Ala Lys Tfp Thr Glu Glu Go Glu Gin Tyr Arg Gin Arg Leu Ala Pro 130 135 140 Go Ala Gin Glu Leu Lys Asp Leu Thr Lys Gin Lys goes Glu Leu Het 145 150 155 160 170

Gin Ala Lys Leu Thr Pro Vai Ala <3iu Glu Vai Arg Asp Arg Leu Arg 165 170 175

Glu Gin Glu Glu Glu Leu Arg Lys Asn Leu Ala Fro Tyr Ser Set Glu

ISO 185 ISO

Leu Arg Gin Lys Leu Ser Gin Lys Leu Glu Glu Ile Arg Glu Arg Sly 195 SOO 205

Ile Fro Gin Ala Ser Glu Tyr Gin Ala Lys Vai Vai. Glu Gin Leu Ser 21Ô 215 220

Leu Arg Glu Lys Het Thr Pro Leu Glu Glu PJte Lys Giu Arg 225 230 235 240

Leu Thr Pro Tyr Ala Glu Asn Leu Lys Asn Arg Leu Ile Asp Leu Leu 245 250 255

Asp Glu Go to Gin. Lys Thr Mst Ala 260

&lt; 210 &gt; 28 &lt; 211 &gt; 264 &lt; 212 &gt; PRT &lt; 213 &gt; Anas platyrhynchos &lt; 400 &gt; 28 171 mi Arg ¥ al Vai 1

Go Leu Leu Leu. Phe Leu Thr Gly Thr 5 10 15

Gin Ma Arg Tyr SD

Ph® Trp Gin His Asp Glu Pro Gin Ala Pro Leu Asp 35 30

Arg Leu Arg Asp35 'Leu Go Asp Go Tyr Leu 6lu Thr Go Lys Ala Ser 40' 15

Gly Lys Asp Ala 50 lie Ala Gin Phe 01u Ala Ser Ala Go Gly Lys Gin 55 60 hm Mp Uu Lys

Lea Ala Mp Mn Leu 1 70

Thr Leu Gly Ala Ala Ala 75 80

Ala Lye Leu htq

Glu Asp Met Ala Pro Tyr Tyr tys Glu Vai Arg GIu 172 85 90 95

Met Trp Leu Lys Asp Thr Glu be Leu Arg Ala Glu Leu Thr Lys Asp 100 105 110 Leu 61u Glu Vai Lys Glu Lys Ile Arg Pro Phe Leu Asp Gin Phe Ser 115 120 135

Ala Lys Trp Thr Glu Glu Leu '31 or Gin Tyr Arg Gin Arg Leu Ala Gold 150 135 HO

Go Ala Giu Glu Leu Lys Glu. Leu Thr Lys Gin Lys Go Glu Leu Met 145 150 155 160

Glft Gin Lys Leu Thr Fro Vl Ala Glu Giu Ala Arg Asp Arg Leu Arg 165 170 175 G.ly His Vai Glu Glu Leu A cg Lys Asn Leu Ala Prg Tyr Ser ftsp Glu 180 '185 190

Leu Arg Gin Lys Leu Ser Gin Lys Leu Glu Glu lie Mg Glu Lys Giy 195 200 205 lie Fro Gin Ala Ala Glu Tyr Gin Ai a Lys Vai Vai Glu Gin Leu Ser 210 215 220

Asa hm Arg Giu Lys Hat Thr Pro Leu Vai Gin Asp Phe Lys Glu Arg 225 230 235 240

Leu Thr Pro Tyr Ale Glu Asn Leu Lys Thr Arg Phe Ile Ser Leu Leu 245 250 255 h &g; Glu Leu Gin Lys Thr Go Ala 260 &lt; 210 &gt; 29 &lt; 211 &gt; 262 173

&lt; 212 &gt; PRT &lt; 213 &gt; Oncorhynchus mykiss &lt; 400 &gt; 29

Met Lys Pbe Leu Ha Leu Ma Leu Thr He Leu Leu Wing Wing Gly Tbr 1 5 10 15 174

Gin Ma Ohe Pro Met <3ln Ma A $ p AiA HO Ser Gin Lee Glu Bis Go 20 25 30

Lys Ala Ala Leu Seri ty ty lie Ala Glu Vai Lys Leu Thr Ala Gin 35 40 45

Arg Ser 11 «Asp Leu Leu Asp Asp Thr Glu Tyr Lys Glu Tyr Lys Ket 50 55 60

Girs I Thr Gla Ser iee Asp Asn Leu Gin Gin Tyr Ala Asp Ala Thr SS 70 U 30

Be Gin Be Leu Wing Pro Tyr Be Glu Ala Phe Gly Thr Gin Leu Thr 35 90 95

Asp Ala Thr Ala Ala Go Arg Ala Glu Va M »t Lys Asp Go Glu Glu 100 105 Π0

Leu Arg Ser Gin Leu Glu Pro Lys Arg Ala Glu Leu Lys Glu Vai Leu 115 120 125

Asp Lys His Ile Asp Glu Tyr Arg Lys Lys Leu Glu Pro Leu lie Lys 130 135 140

Glu His 11 «Glu Leu Arg Arg Thr Glu Hei Gly Ala Phe Arg Ala Lys 1« 150 155 160

Hat Glu Prc Ile Go Glu Glu Leu Arg Wing Lys Go Wing Ile Asa V'ai 165 170 175

Glu Glu Thr Lys Thr Lys Leu Met Prole Go Glu Ile V Al Arg Ala 160 185 190

Lys Leu Thr Glu Arg Leu Glu Glu Leu Arg Thr Leu Ala Ala Pro Tyr 195 200 205

Ala Glu Glu Tyr Lys Glu rank Met Ile Lys Ala Go Gly Glu Vai Arg 210 '215 220

Glu Lys Will Be Pro Leu Be Glu Asp Phe Lys Gly Gin Go Gly Pro 225 230 235 240 175

Ala Ala Glu Gin. Ays Lys Gin Lys L &amp; Leu Ala Pha ryr Gltt Thr Ile 245 2SQ 255

Be Gin Wing Het Lys Ma 260

&lt; 210 &gt; 30 &lt; 211 &gt; 262 &lt; 212 &gt; PRT &lt; 213 &gt; Salmo trutta &lt; 400 &gt; 30 176 ttet Lys Phe Leu Ala Leu Ala Leu Thr He Leu Leu Ala Ale Al Thr 15 5 15

Gin Ala Vâl Pro Met 61 n Ais Asp Ala Ptq Sar Gin Leu Glu His Vai 20 25 30

Lys Goes Ala Hat Met 61 u Tyr Hat Ala Gin Goes Lys Glu Thr Gly Gin. 35 40 45

Arg Set II »Asp Leu Leu Asp Asp Thr 61 u Phe Lys Glu Tyr Lys Go 50 55 € 0

Gin Leu Ser Gin Gin Leu Asp Asn Leu Gin Gin Tyr Ala Gin Thr Thr 65 70 75 80

Ser Gin Ser Leu .Ala Hro Tyr Ser Glu Ala Phe Gly Ais Gin Leu Thr 85 90 SS

Asp Ala Ala Ala Ala Vai Atg Ala Glu Vai Met Lys Asp Vai Glu Asp 100 105 lio

Go Arg Thr Gin Lett Glu $? Ro Lys Arg Ala Glu Leu Lys Glu Vai Leu 115 120 125

Asp Lys His He Asp Glu Tyr Arg Lys Lys Leu Glu Pro Leu He Lys 130 135 140

Glu He Glu Glu Gin Arg Arg Thr Glu Leu Glu Ala Fhe Arg Val Lys 145 150 155 '160

Met Glu Pro Will Go Glu Giu Met Arg Ala Lys Will Be Thr Asn Go 165 170 '17S 177

Glu Glu Thr Lys Ala Lys Lm mt Pro He Vai Glu Thr goes Arg Ma 180 185 190

Lys Leu Thr Glu Arg hm Glu Glu Leu Arg Thr Leu Ala Ala Pro Tyr 195 200 205

Ala Glu Glu Tyr Lys Glu Glu Hat Ptw Lys Ma Go Gly Glu Go Arg 210 215 22G

Glu Lys Go Gly Pro Leu Thr Asn Asp Phe Lys Gly Gin Go Gly Pro 225 J 230 235 240

Ala Alâ Glu &lt; * Ala Lys Glu Lys leu Mst Asp Phe Tyr Glu Thr Ile 245 250 255

Ser Gin Ala Met Lys Ala 260 &lt; 210 &gt; 31 &lt; 211 &gt; 258 &lt; 212 &gt; PRT &lt; 213 &gt; Salmo salar &lt; 4 0 0 &gt; 31 178

Set Lys Phe Leu Vai Leu Ala Lee Tíir lie Leu Leu Ala Ala Gly Thr 1 5 10 15

Gin Wing Phe Pro Met Gin Ala Asp Ala Prô Ser Gin Leu Glu Eis Go 20 '25 30

Lys Ala Wing Leu Asn Met Tyr lie Ala Gin Go Lys Leu Tht Ala Gin 35 40 45

Arg Sér He Asp Leu Leu Asp Asp fhír Glu Tyr Ly $ GlU Tyr Lys Set 50 55 60

Gin Leu Ser Gin Ser Leu Asp Asn Leu Gin Gin Phe Ala A & p Ser Thr 65 70 75 80

Ser Lys Ser Irp Pro Pro Thr Pro Arg Ser Ser Ala Pro Ser Cys Asp 85 9Q 9.5 179

Ala Thr Ala Thr Vai Arg Ala Glu Va Mefc Lys Asp Va Glu Asp Go 100 105 110

Arg Thr Gin Leu Glu Pro Lys Arg Ma Glu Leu Thr Glu Vai Leu Asn 115 120 125

Lys Kis Ile Asp Glu fyr Arg Lys Lys Leu Glu Pro Leu lie Lys Gin 130 135 140

His Ile Glu Leu Arg Arg Thr Glu Met Asp Ala Phe Arg Ma Lys Xle 145 159 15 $ 160

Asp Pro Vai Vai Glu Glu Met Arg Ala Lys Will Ala Go Asn Go Glu 165 'Π0 175

Glu Thr Lys Thr Lys Leu Met Prole Val Glu Ile Va.1 Arg Ala Lys ISO 185 190

Leu Thr Glu Arg Leu Glu Glu Leu Arg Thr Leu Ala Ala Prô Tyr Ala 135 390 305

Glu Glu fyr Lys Glu Glu Met Ph.e Lvs Ala Vai Giy Glu Vai Arg Glu 210 215 220

Lyg Go Wing Psc Lou Be Glu Asp Phe Lys Ala Arg Trp Ala Pro Pro 225 230 233 240

Pro Arg Arg Pro Ser Lys Ser Ser Trp Leu Ser Thr Arg Pro Ser Ala 245 250 255

Arg Pro

&lt; 210 &gt; 32 &lt; 211 &gt; 262 &lt; 212 &gt; PRT &lt; 213 &gt; Brachydanio rerio 180 &lt; 400 &gt; 32

Met Lys Pha Goes Ala Ma Lao fhr Lau Lao Lm. Ala Leu Gly Ser

IS I IS

Gii Ala Asr $ hm Ph «Glo Ala &amp; P Ala Pro Thr G.ia Lao GI« Hl Tyr 20 25 30 181

Lys Ma Ala Lea Vai Tyr Leu Asn Gin Go Lys Asp Gin Ala Glu 35 40 * 45

Lys Ala Leu Asp Asm Leu Asp Gly Thr Asp Tyr Glu Gin Tyr Lys Leu 50 55 60

Gin Leu Ser Glu Ser Leu Thr Lys Leu Gin Glu fyr Ma Gin Thr Thr 65 70 75 SG

Ser Gin Ala Leu Thr Pro Tyr Ala Glu Thr He Ser Thr Gin lea Met 85 90 95

Glu Asn Thr Lys Gin Leu Arg Glu Arg Vai Met Thr Asp Vlu Glu Asp 100 .105 Π0

Leu Arg Ser Lys Leu Glu Pro His Arg Ala Glu Leu Tyr Thr Ala Leu 115 120 125

Gin Lys Bis Ilô Asp Glu Tyr Arô Glu Lys Lôu Gltt P. &gt; Go Phô Gin 130 135 140

Glu Tyr Ser Ala Leu Asn Arg Gin Asn Ala Glu Gin Leu Arg Ala Lys 145 ISO 155 160

Leu Gl «Pro Leu Met Asp Asp 11« Arg Lys Ala Phe Glu Ser Asn Ile 165 170 175

Glu Glu Thr Lys Ser Lys Will Go Pro Met Will Glu Ala Go Arg Thr 180 185 190

Lys Leu Thr Glu Arg Leu Glu Asp Leu Arg Thr Met Ala Ala Pro Tyr 195 '200 205

Ala Glu Glu Tyr Lys Glu Gin Leu Vai Lys Ala Go Glu Glu Ala Arg 210 2X5 220

Glu Lys lie Ala Pro Bis Thr Gin Asp Leu Gin Thr Arg Met Glu Pro 225 230 235 '240

Tyr Met Glu Asn Go Arg Thr Thr Phe Ala Glti Hefc Tyr Glu Thr Ile 245 250 255 182 Ala Lys Ala Ile Gin Ala &lt; 210 &gt; &Lt; 211 &gt; 260 &lt; 212 &gt; PRT &lt; 213 &gt; Sparus aurata &lt; 400 &gt; 33 183

Met Lys Pfee Ala Ala Leu Ala Leu Ala Leu Leu Leu Ala Go Gly Ser 1 5 10 15

His Allah Wing Ssr Het Gin Wing Asp Wing Pro Ser Gin Leu Asp Kls Wing 20 1 25 30

Ala Vai Leu Asp Go 35

Leu Thr Gin Vai Lys Asp Met Ser Leu 40 45

Thr

Arg Ala Go Asm 6.1 «Leu Asp Asp Pro Gin Tyr AXa Glu SC SS 60

Asn leu Ala Gin Arg Ile Gia Glu Met Tyr Thr Gin Ile lys Thr Leu 65 70 75 80

Gin Gly Will Be Pro mt Thr Asp Be Phe Tyr Asa Thr Will Hat

Glu Vai Thr Lys Asp Thr Arg G.l Ser Leu Asn Val Asp Leu Glu Ala 100 105 110

Leu Lya Ser Ser Leu Wing Pro Gin Asn Glu Gin Leu Lys Gin Go Ile US 12Õ 125

Glu Lys Bis Leu. Asn Asp Tyr Arg Thr Leu Leu Thr Pro Ile Tyr Asn 135 140

Asp Tyr Lys Thr Lys His Asp Glu Glu Met Ma Ala Leu Lys Thr Arg 145 150 155 160

Leu Glu Pro Go Hfet Glu Glu Leu Arg Thr Lys Ile Gin Ala Asn Go 165 170 175

Glu Glu Thr Lys Ala will Leu Met Pro 'et Vai Glu Thr Vai Arg Thr 184 ISO 18 S 130 tys Vai Thr Gle Arg Leu Çiu Ser Leu & Glu Go Vai Gin Fro Tyr 195 200 205

Go Gin Glti Tyr Ly $ Glu Gin Hei Ly Gin Gin Tyr &amp; p Gin Gin 210 215 * 220

Thr value Mp Thr Mp Ma Uu Mq Thr iys Π® Thr Wm Leu Vai Gin 22S 230 235 240

Glo Jla Lys Go Lys Met Asai Ais lie Phe Glu 11® Ile Ala Ser Ser 245 250 255

Go Thr Lys Ser 260

&lt; 210 &gt; &Lt; tb &gt; 396 &lt; 212 &gt; PRT &lt; 213 &gt; Homo sapiens &lt; 400 &gt; 34 185

Mst Phe Leu Lys Ala Goes Leu Leu Ma Leu Goes Ala Goes Ala 1 5 10 15 61 y Ala ftrg Ala Giu Goes Ala Asp Gin Goes Ala Tht Go Met ϊχρ 2Q 25 38

Asp Tyr Phe Ser Gin Leu Set Ass Aso Ala Lys Glu Aya Go Glu His 3S 40 45

Leu Gin Lys Be Giu Leu Thr Gin Gin Leu ftsn Ala. Leu Ph «Gin Asp 50 5-5 60

Lys Leu Glv Glu Go Asn Thr Tyr Ala 61 and Asp Leu. Gin Lys Lys Leu 65 70 75 80

Go to Phe Ala Thr Glu Leu Kis Glu Arg Leu Ala Lys Asp Ser Glu 85 90 '§S

Lys Leu Lys Giu Glu Xle Gly Lys Glu Leu Glu Glu Leu Arg Ala Arg IDO 105 3.10 186

Leu Leu Fro Ris Ala Asn Glu Will Be Slrt Lys Ile Gly Aap Asn hm 115 120 125

Arg Glu Leu Glu Glu Arg Leu Glu Pt or Tyr Ala Asp Gin Leu Arg Thr 130 135 140

Gin Vai Ass Thr Gla Ala Glu Glu Leu Arrr A.rgr Glu Leu Thr Prss Tyr 145 150 155 160

Ala Glu Arg Sfet Glu Arg Vai Leu Arg Glu Asn Ala Asp Ser Lea Gin 165 &quot; 170 175

Ala Set Leu Arg Pre His Wing Asp Glu Leu Lys Wing Lys Ile Asp Gin ISO 185 190

Asa Go Glu Glu Leu Lys Gly Arg Leu Thr Pro Tyc Ala Asp · Glu Phe 1S5 20Q 205

Lys Go. Lys Ile Mp Gin Thr Go Glu Glu Leu Arg Arg Set Leu Ala 210 '215 220

Pto Tyr Ala Gin Asp Thr Gin Glu Lys Leu Aan Ris Gin Leu Glu Gly 225 230 235 240

Leu Thr Fhe Gin Rer. Lys Lys Aso Ala Glu Glu Leu Lys Ala Arg lie 245 250 255

Be Wing Be Wing Glu Glu Leu Arg Gin Arg Wing Wing Pto Leu Wing Glu 260 265 270

Asp Go Arg Gly Asn Leu Lys Gly Asa Thr Glu Gly Leu Gin Lys Ser 275 280 285

Leu Wing Glu Leu Gly Gly His Leu Asp Gin Glu Glu Glu Phs Arg 230 235 300

Arg Alo Go Glu? Rc Tyr Gly Glu Asm FM Mn Lys Ala Leu vai Gin 305 '310 313 320

Gin Met Glu Gin Leu Arg Gin Lys Leu Gly Pto His Wing Gly Asp Go 325.330 335 187

Slu Gly Bis Leu Ser Ph «Leu Glu Lys Asp Leu Arg Asp Lys Go Asn 340 345 350 187 Ser Phe Phe Ser Thr 355

Poè Lys Glu Lys 360

Glu Ser Gin Asp Lys 365

Thr Leu

Ser Leu Pro Glu

Leu Glu Gin Gin Gin

Glu Gin Gin Gin Glu Gin Gin 370 375 380

Gin Glu Gin Vai Gin ^ et Leu Ala Pro Leu Glu Ser 385 390 395 &lt; 210 &gt; &Lt; tb &gt; 429 &lt; 212 &gt; PRT &lt; 213 &gt; Macaca sp. &lt; 400 &gt; 35 188

Mefc Phe Leu Lys Ala Will Go Leu Thr Leu Will Lea Will Go Ala Will Thr 1 S 10 15

Gly Ala Arg Ala Glu Will Be Ala Asp Gin Will Ala Thr Will Ket Trp 20 25 30

Asp Tyr Phe Ser Gin Leu Ser Ser Asa Wing Lys Gin Wing Go. Gin His 35 40 45

Leu 61 n Lys Ser Gin Leu Thr Gin Gin Leu Asn Lea Leu 6he Gin Asp 50 55 60

Lys Leu Gly Glu Go Asn Thr Tyr Ala Gly Asp Leu Gin Lys Lys Leu 65 70 &quot; 75 80

Go Pro Phe Ala Thr çiu Leu His Glu Arg Leu Ala Lys Asp Ser 61 «85 90 SS

Lys Leu Lys Glu Glu Xle Arg Lys Glu Leu Glu Glu Vai Arg Ala Arg 100 105 11.0

Leu Leu Pro His Wing Asn Glu Will Be Gin Lys Ile Gly Glu Asn Go 115 115 125

Arg Glu Leu 61 to Gin Arg Lee Gin Pro Tyr Thr Asp Gin Leu Arg Thr 130 135 140 189

Gin Vai Aci Thr GiU Gin H5 ISO Ala Gin Arg Glu Arg Vai Leu 165 Tb * Ser Leu Arg Fr o Is Ala Asp 180 Asn Go Glu Glu Leu Lys Glu Arg 195 200 Lys Go Lys Ile Asp Gin Thr Go 210 215 FfO Tyr Ala. Glu Asp Ala Cl »Glu .225 230 Leu Ala Fhe Glrs Mst. Lys Lys Asn 245 Be Ala Ser Ala Glu Glu Leu Arg 260 Asp Met Arg Gly Asn Leu Arg Gly 275 280 Leu Ala Glu Leu Gly Gly His Leu 250 295 Leu Arg Go Glu Pro Tyr Gly Glu 305 31G 61 n «et Glu Glrs Leu 325 Arg Gin Lys Glu Gly Bis Leu Ser Phe Leu Glu 340 Ser Phe Phe Ser Thr Phê Lys Glu 353 360

Leu Arg Arg 610 Leu Thr pro Tyr 15 S 160 Arg Glu Aso Ala Asp Ser Leu Gin 170 175 Gin Leu Lys Ala Lys lie Asp Gin 185 190 Leu Thr Pro Tyr Ala Asp Glu Phe 205 Gi u Glu Leu Arg Arg Ser Leu Ala 220 Lys Leu Asn His Gin Leu Glu Gly 235 240 Ala Glu Glu Leu Lys Ala Arg Ile 250 255 Gin Arg Leu Ala Pro Leu Ala Glu 265 270 Asn Thr Glu Gly Leu Gin Lys Ser 285 Asp Arg Ris Vsl Glu 61 s Phè Arg 300 Asn Phe Asn Lys Ala. Leu Vai Gin 315 320 Leu. Gly Pro His Wing Gly Asp Go! 330 335 Lys Asp Leu Arg Asp Lys Go Asn 345 35S Lys Glu Oer Gin Asp Aso Thr Leu 365 190

Ser Leu Pro Glu Pro Glu Gin Gin Arg Gin Gin, Gin Gin Glu Gin Gin 370 '375' 380

Gin Glu Gin Glu Gin. Glu Gin Gin Gin Gin Gin Gin Gin Gin Gin 385 390 335 400

Glu Glu Gin Gin Arg 61u Gin Gin Gin Gin Glu Gin Gin Gin Glu Gin 405 410 415

Gin Gin Glu 6l.n Go Gin Met Allo Ala Pró Léu Glu Sul 420 425 &lt; 210 &gt; 36 &lt; 211 &gt; 395 &lt; 212 &gt; PRT &lt; 213 &gt; Mus sp. &lt; 400 &gt; 36 191

Met Phe Leu Lys Ma Ala Go Leu Thr Leu Ma Leu Go Ma 11a Thr

1s 10 IS

Gly Thr Arg Ala Glu Go Thr Ser Asp Gin Go Ala Asn Go Go Irp 20 25 30 ftsp Tyr Phe Thr Gin Leu Ser Asn Asn Ala Lys Glu Ala Go Glu Gin 35 40 45

Fhe çln Lys Thr Asp VaX Thr Gin Gin Leu Ser Thr Leu Phe Gin Asp S0 55 60 lys Leu Gly Asp Ais Ser Thr Tyr Ala Asp Gly Vai His Wing Lys Leu 65 10 75 80

Go Pro Ph «Go Go Go Go Go Go Go Go

Arg Val Lys Glu Glu Ile Lys Lys Glu Leu Glu Asp Leu Arg Asp Arg 100 105 lio ttet Het Pr o His Ala Asn Lys Go Thr Gin Thr Phe Gly Glu Asm Het 115 120 125

Gin Lys Leu Gin Glu His Leu Lys Pro Tyr Ala Go Asp leu Gin Asp 192 130 133 140

Gin lie Asa Thr Gin Thr Gin Glu «et Lys tu Gin tu Thr Pro Tyr

145 150 .155 ISO J.Iíí Gin árg mt Gin Thr Thr Xle Lys Glu Asn VeX Asp Asa Lea Hí * 165 Π0 175

Thr Ser Mefc Met Fro? I Ma Thr Asn L * u Lys Asp tys Phe Asts Arg 180 165 190

Asn H * t Glu Glu Leu Lys Gly Hxs Leu Thr Pro Arg Ala Asn Glu Leu 195 200 205 ·

Lyn Ala Thr Xis Asp Gin Mn Leu Glu A p Leu Arg Arg Ser Leu Ala 210 215 220

Pro-U Thr Go Siy Go Gin Glu LyS Leu A $ His Gin «et Glu Gly 225 230 235 240

Leu Ala The Gin «et Lys Lys Asn Ala Si a. Glu Leu Gin Thr Lys Go 245 250 255

Be Wing Lys lie. Asp Gin Leu Gin Lys Wing Leu Wing Leu Vai Glu 260 265 270

Asp goes Gin Ser Lys Vei Lys Gly Asn Thr G 1u Gly Leu Gin Lys Ser 275 2S0 205

Leu Glu Asp Leu Asn Arg Gin Leu Glu Gin Gin Go Glu Glu Phe Arg 250 295 300

Arg Thr Go Glu Pro Met Gly Glu Met Phe Asn Lys AU Leu ¥ sl Gin 305 310 315 320

Gin Leu Glu Gin Phe Arg Gin Gin Leu Gly Pro Asn Ser Gly Glu Go 325 330 335

Glu Ser His Leu Ser Phe Leu Glu Lys Ser Leu Arg Glu Lys Go Asn 340 345 &quot; 350 Ser Phé Het Ser Thr Leu Glu Lys Lys Gly be Pró ASP Slft Fro Gin 355 3ê0 365 193 193

Ais Leu Pro Leu Fro Glu Mn Ma Gin Glu Gin Ma Gin Glu Mn Λ 370 375 38G

Gin Glu Gin Go Pro Gin Lys Pro Leu Glu Ser 385 390 395 &lt; 210 &gt; 37 &lt; 211 &gt; 401 &lt; 212 &gt; PRT &lt; 213 &gt; Baboon sp. &lt; 400 &gt; 37 194

Gly Ala Arg Ma G.iu Will Be Ala Aap Gin Go Ala Thr Go to jffet? Rp 1 3 '10 15

Asp Tyr Phac Ser Gin read Ser Ser Asn Ala Lys Glu Ala Go Giu His 20 25 30

Leu Gin Lys Ser Glu read Gin Gin Leu Aan Ala Leu. PM Gin Asp 35 40 '45 lys Leu Gly Glu Go Asa Thr Tyr Ala Gly Asp Leu Gin Lys Lys Leu 50 55 10

Go Pro Phe Ala Thr Glu hm His Glu Arg Leu Ala Lys Mp Ser Lys 65 70 75 80

Lys Leu Lys Glu Glu Ile Arg Lys Glu Leu Glu Glu Go Arg Arg to Arg 85 90 95

Leu Leu Pro His Wing Wing Glu VaX Ser Gin Lys lie Gly Glu A $ n Go 100 105 '110

Arg Glu Leu Gin Gin Arg Leu Glu Pro Tyr Thr Asp Gin Leu Arg Thr 115 120 125

Gin Go Asa Thr Gin Thr Glu Gin Leu Arg Arg Gin Leu Thr Pru Tyr 130 135 '140.

Ala Glu Arg Hèt Glu Arg Vai Leu Arg Glu Asn Ala Asp Ser Leu Gin 145 ISO 155 160 195

Tftr Shr Lean Arg Pro His Wing Asp Gin. Lys .Lys Ma lys lie Asp Gin 165 170 175

Asn valuu çiu Lea Lys Gly Arg Leu m P * o Tyr Ala a P p Glu? He 180 185 190

Lys Val Lys Xle Asp Gin Thr Val Glu Glu Leu Arg Arg Ser Leu Ala 195 290 205

Pro Tys Ala Gin Asp Ala Gin Glu Lys Leu Asn 8is ol Leu Glu Gly 210 215 &quot; 220

Leu Wing Phe Sln Met Lys Lys Asn Ala Glu Glu Leu Lys Ala Arg He 225 230 235 240

Be Wing To Be Wing. Glu Glu Leu Arg Gin Arg Leu Ala Pro Leu Glu rank 245 250 255 ASP Ket Arg Gly Aârt Leu Arg Gly Asn. Thr Glu Gly LsU Glfi lys Ser 26Q '265 270

Leu Ala Glu Leu Gly Gly His U »Asp Arg His Val Glu Glu Phe Arg 275 2S0 285

Leu Arg Val »Pro Tyr Gly Glu Asn Phe Asn Lys Ala Leu val Gin 23Õ 295 300

Gin Mfit Glu GXn Leu Arg Gin Lys Leu Gly Pro His Ala Gly Asp va'1 305 310 315 3.20

Glu Gly Hii

Read

Ser Phe Leu Glu Lys Asp Leu Arg Asp Lys Go Asn 325 "330 &quot; 335

Ser Phe Phe Ser Thx PAe Lys Glu Lys Glu Ser Glft Asp Asp Thr L U 340 345 359

Ser Leu Pro Glu Pra Glu Gin Gin Glu Gin Gin Gin Glu Gl.n Glu 355 360 365

Gin Gin Gin Gin Gin Gin Gin Glu Gin Gin Gin Gin Glu Gin Gin 370 375 380

Gly Gin. Glu Gin Gin Gin Glu Gin Go Gin Met Leu Ais Pro loa Glu 196 385 390 395 400

&Lt; 210 &gt; 38 &lt; 211 &gt; 382 &lt; 212 &gt; PRT &lt; 213 &gt; Its sp. &lt; 400 &gt; 38 197 ístet Pfte Leu Lys Ma Go Go Leu Be thy Wing Leu Go Ala Go Thr 1 5 10 15

Gly Ala Arg Ala Glu Go Asa Ala Asp Gin Go Ala Thr Go Set Trp 20 25 ^ 30

Asp Tyr Ph © Ser Gin Leu Gly Ser Asn Ala Lys Lys Ala Vai Slu His 35 49 45

Leu Gin Lys Ser Glu Leu Thr Gin Gin Leu Asn Thr Lea Phe Gin Asp SG 55 60

Lys Leu Giy Glu Go Asn Thr Tyr Thr Glu Asp Leu Gin Lys Lys Leu € 5 70 '75 80

Go Pro Phe Ala Thr Glu Leu His Glu Ara Thu Lys Asp Ser Glu 85 90 95

Lys Leu Lys Glu Glu He Arg Arg Glu Leu Glu Glu Leu A.rg

Leu Leu Pr © Eis Ala Thr Glu Will Be Glu Lys ile Gly Asp Asa Vai 115 120 125

Arg Glu Leu Gin Glu Arg Leu Giy Pro Phe Thr Gly Gly Leu Arg Thr 130 '135 140

Glu Vai Asn Thr Gin Go Gin Gin Leu Gin Arg Gin Leu Lys Pro Tyr 145 150 155 160

Ala Glu Arg fet Glu Ser Vai Leu Arg Gin Asn lie Arg Asn Leu Glu X 5 170 175 198

Ala Ser Go Pro Wing Tyr Ala Asp Glu Phl Lys Ala Lys He Asp Qln 180 185

Asn Go to Glu Leu Lys Gly Ser Leu Thr Pro Tyr Ala <3lu çlu Leu.

Lys Wing Lys He Asp Gin Asn Go Glu Glu Leu Arg Arg Sir Leu Alâ 210 '2X5 220

Pro Tyr Ala Gin Asp Go Gin Glu Lys Leu Asn His Gln Leu Glu Gly 225 '230 235 240

Leu Wing Phe Gin Met Lys Lys Gin Wing Glu Glu Leu lys Wing Lys Ile 245 250 255

Be Ala Asn Ala Asp Glu Leu Arg Gin Lys Leu Go Pro Go Ala Glu 2ÊQ '2 € 5 * 270

Asn Go His His His His Leu lys Gly As "Tfcr Glu Gly Leu Gin Lys Ser 275 2SG 205

Leu Leu Glu, Leu Arg Ser His Leu Asp Gin Gin, Vai Glu Glu Phe Arg 290 295 300

Leu Lys Go Glu Pro Tyr Gly Glu Thr Phe Asn Lys Ala Leu Vai Gin 305 .310 '315' 320

Gin Vai Glu Asp Leu Arg Gin Lys Leu Gly Pro Leu Ala Gly Asp Vai 325 "330 335

Glu Gly His Leu Ser Phe Leu Glu Lys Asp Leu Arg Asp Lys Go Asn 340 345 * 350

Thr Phe Phe Ser Thr Leu Lys Glu Glu Ala Ser Gin Gly Gin Ser Gin 355 360 365

Ala Leu Pro Ala Gin Glu Lys Ala Glu Ala Pro Leu Glu Gly 370 375 380 199 &lt; 210 &gt; 39 &lt; 211 &gt; 391 &lt; 212 &gt; PRT &lt; 213 &gt; Rattus sp &lt; 4 0 0 &gt; 39 200

Phe Leu Lys Ala Leu Leu Thr Leu Leu Leu Leu Ala Ile Thr 1 5 10 IS Gly Thr Gin Ala Glu Will Thr: Ser Asp Gin Will Ala Asn Will mt Trp 20 25 30 Asp Ty.r Phe Thr Gin Leu 3ec Asn Asn Ala Lys Glu Ala Go Giu Gin 35 40 V 45 Leu Gin Lys Thr Asp Go Thr Gin Gin Leu Asn Thr Leu Phs Gin Asp 90 $ 5 60 Lys Leu Gly Asn n & Asn Thr Tyr Ala Asp Asp Leu Gin Asn Lys Lee. 65 70 75 80 Go Pro Phe Ala Go Gin Luu Be Gly His Leu Thr Lys Giu Thr G.lU 85 90 95 Mg goes Arg Glu Glu Ile Gin Lys Glu Leu Glu Asp Leu Arg Wing Asn 100 105 in Met Met Pro His Wing Asn Lys Will Be Gin Met Phe Gly Asp Asn Go 1.15 120 125 Gin Lys Leu Gin Gi His Leu Mq Pro Tyr Ala Thr Asp Leu Gin Ala 130 135 HO Gin lie Ala Gin Tft * Gin Asp Met Lys Arq Gin Leu thr Pro Tyr 145 150 155 160 11 and Gin Arg Met Gin Thr Thr Ile Gin Asp Asn Go Glu Asn Leu Gin 165 170 175 S * r Be mt Pro Go PM Ala Asn Glu Leu Lys Gls Lys PM Asn Glfi 180 185 190 Asn Met Glu Gly Leu Lys Gly Gin Leu Thr Pro ftrg Ala Asn Glu Leu 135 200 205 Lys Aiâ "¢ He Asp Gin Asn Leu Glu Asp Leu Arg Ser Arg Leu Ala &gt; 10 215 220 201

Pro Leu Ala Glu Gly Vai Gin Glu Lys Leu Asa HIs Gla Môt GXtt Gly 225 330 235 240

Leu Wing Phe Gin Met Lys Lys Ala Glu Slu Leu Glu Thr Lys Vai 245 '250 255

Ser Thr Μη Ile Asp Gin Leu Gin Lys Aan Leu .Ala? Ro Leu Vai Glu 260 .265 270

Âap VÂ · Glu Ser Lys Leu Lys &lt; 51 and Aso Thr Glu Gly Lm Gin Lys Ser 275 280 28S

Leu Glu Asp Leu Aan Lys Gin Leu Asp Gin. Gin. Vâl Glu Vai Phe ftrg 29U 2.95 300

Arg Ala, Go Glu. Pro Leu Gly Asp Lys Phe Asn het Ala Leu Vai Gin. 305 310 315 320

Gin Met Glu Lys Phe Arg Gin Glu Leu Gly Ser Asp Ser Gly Asp Go 325 330 335

Glu Ser is Leu Ser Phe Leu Glu Lys As Leu Arg Glu Lys Will Be 340 345 350

Ser Phe het Ser Thr Leu Gin Lys Lys Gly Ser Pro Asp Gin Pro tu 355 360 365

Alu Leu Pro Leu Pro Glu Gin Go Gin Glu Gin Go Gin Glu sin Go 370 375 38Ô

Gin Pro Lys Pro Leu Glu Ser 385 300 202

REFERENCES REFERRED TO IN THE DESCRIPTION The present list of references cited by the applicant is presented merely for the convenience of the reader. It is not part of the European patent. Although care has been taken during the compilation of references, it is not possible to exclude the existence of errors or omissions, for which the IEP assumes no responsibility.

WO 9312143 A, SIRTORI • US 5643757 A, SHA-IL • US 5990081 A, AGELAND • WO 9637608 A, RHONE-POULENCE ROHRER • WO 9012879 A, Sirtori • WO 9413819 A • WO 9856906 A • WO 9531540 A, HOPPE [0083] • EP 0036776 A, Goeddel WO 9418227 A WO 9946283 A [0149] US 5922680 A [0149] A U.S. 5,780,434 A [0149] A U.S. [1549] [15] [15] [15] [15] [15] [15] [15] [15] [15]

Literature quoted in the description, in addition to the patents of invention • Braschi et al. J Lipid Res 1999, vol. 40, 522-532 [0005] • Braschi et al. Biochemistry, 2000, vol. 39, 5441-5449 Glass et al. J Biol Chem, 1983, vol. 258, 7161-7167 Sorensen et al. Gene, 1995, vol. 152, 243-245 [0035] Miyazaki et al. Arteriosclerosis, Thrombosis, and Vascular Biology, 1995, vol. 15, 1882-1888 Sha PK et al. Circulation, 2001, vol. 103, 3047-3050 [Nielsen et al. FEBS Lett, 1997, vol. 412, 388-396 Lorentsen et al. Biochem J, 2000, vol. 347, 83-87 Nielsen BR; Kastrup JS; Rasmussen H; Holtet TL; Graversen JH; Etzerodt M; Thogersen HC; Larsen IK. FEBS-Letter, 1997, vol. 412, 388-396 Pack P .; Plückthun, A. Biochemistry, 1992, vol.31, 1579-1584 [Muller, K. M .; Arndt, K. M .; Alber, T. Meth. Enzymology, 2000, vol. 328, 261-281 Tissue Culture, 1973 [0129] Ellgaard L. et al. Eur. J. Biochem., 1997, vol. 244 (2), 544-51 [0154] Bergeron J. et al. Biochem. Biophys. Acta, 1997, vol. 1344, 139-152 Kozyraki R; Fyfe J; Kristiansen M; Gerdes C;

Jacobsen C; Cui S et al. The intrinsic factor-vitamin B12 receptor, cubilin, is a high-affinity apolipoprotein A-I receptor facilitating endocytosis of high-density lipoprotein. Nat Med, 1999, vol. 5 (6), 656-661 [0168] 204 • Lorentsen RH; Graversen JH et al. Biochemical Journal, 2000, vol. 347, 83-87 [0177] • Holtet et al. Protein Science, 1997 [0179] • Lorentsen et al. Bíochem. Journal, 2000 [Nielsen et al. FEBS, 1997 [0179] • Nielsen et al. Acta Cryst D., 2000 [0179] Bergeron et al. Biochem Biophys Acta, 1997, vol. 1344, 139-152 [0193] • Bolivar et al. Gene, 1977, vol.2, 95 [0193] • Chang et al. Nature, 1978, vol. 275, 617-624 [0193] • Ellgaard et al. Dissection of the domain architecture of the a2macroglobulin-receptor-associated protein. Eur J Biochem, 1997, vol. 244, 544-551 [0193] • Fiers et al. Nature, 1978, vol. 273, 113 [0193] • Goeddel et al. Nature, 1979, vol.281, 544 [0193] • Hess et al. Advances in Enzyme Regulation, 1969, vol. 7, 149-166 [0193] • Hitzman et al. Journal of Biological Chemistry, 1980, vol. 25, 12073-12080 [0193] • Holland et al. Biochemistry, 1978, vol. 17, 4900 [0193] • Holtet, T.L .; Graversen, J.H .; Thogersen, H.C .; Etzerodt, M. Domains and shared motifs in plasminogen ligand interaction. 21st Century Annual Lorne Conference on Protein Structure and Function, held Melbourne, Australia, 04 February 1996 [0193] • Itakura et al. Science, 1977, vol. 198, 1056 [0193] Jones. Genetics, 1977, vol. 85, 23-33 [0193] • Kastrup, J.S. Lecture at Minisymposium held by EU HCM contract CHRX-CT93-0143: Protein Crystallography I in Hamburg, 13 December 1996 [0193] • Kingsman et al. Gene, 1979, 141 [0193] 205 Larsen, I.K .; Nielsen, B.B .; Rasmussen, H .; Kastrup J.S, Poster, 17th International Crystallography Congress, 08 August 1996 [0193] • Neame, P.J .; Boynton, R.E. Protein Soc. Symposium.

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Lisbon, 12/15/2010

Claims (23)

An apolipoprotein construct for use as a medicament having the general formula apo-AX, wherein apo-A is an apolipoprotein component selected from the group consisting of apolipoprotein AI, apolipoprotein AII, apolipoprotein AIV, an analog or variant thereof which shares at least 70% sequence identity to a sequence selected from the group consisting of SEQ ID NO 15: E &amp; ãavitiav LOtfesaRRH fhqqdeffqs mdrvkdlm vYvavtKtsG mvtsQmGs only ilSKBL 'rSKhEEíLGP YTQEFSímB KET1GLR8M SKDLESVKM 120 VQFYLDDFQK KHQ8EHELYK QKVEPLRfiEl QEQMtQKLBS iQSKLSPWSS Β8Β8Μ' βν 180 eMRTHtftPlt SDElRQRLM RLEALKSNGG ARLAEYHAKA TEKLSTtSM MPH. & PLRQ 240 GLLPVLESFK. VSFISALEEy TKKLNTQ 2 &amp; Ί SEQ ID NO 16: HKftAVlTLAV tFLTGSQÃM FWQQDEPBQS PWDRVKDMT VYVDfBL8BSG RtHVSQFKGS 60 ttSBBUnaSt LD »O ¥ TSI FSKUEQLGP VTQEFSTDNU KETEt-tEQEH SKDtEEVKM 120 VOPYLDDFÇK KWQEEMELYR QKVSPLRM1 QEGAHQK &amp; HS LQFJÍLSPLGS EMRDRARAHV 180 DWaiatASÍ SDBMBRLAft RLEÂLKE8GG AOLASYHAKA ÍESLSTLSEK AKPM.E.D1SQ 240 Î »Î» Î »K &amp; LE & TGSQftRí FWQQDEPPOT FWDRVKSLVT YYVEBLKDSG HHVSQFE6S 60 ΗΛΚΒΗΙΙΚΕ. IDSWÓSnSY VSI &amp; BEOIfiP VtQElííóKIíS KETEGLRQEM SKDLEEVKAK 120 fQPYlDBFOK XHfflSfâffim OCTEPLRAEL · HESTRQJS.il IBESUFU 'EVRDRARAHV 18 BftLRmftPY SDEMSRm RUAIKEBSS &amp; KL &amp; EYHMA SSHLSTLSEK MRALEDLRQ 240 GÍíLPVL &amp; SFK VSfLSMiEEY TKSLSTQ 267 SEQ ID NO 18: MKAWLTLRY LFLTSSQRM FKQQDBPQSS TORVKDFm YVEMKDSGR fiWAQmSA 60 LSTOim DMDTLASTL Stasi SKVREQLGFV TQEFWDNLE-K ^ QBHS KDXíEÊVKQKV 120 QPYLDSFQKK KHEBVEIYRQ KVAPIGKSFR GABQKV £ $ 160 LBDRARAitVE EL QDKLSPLftQE TMQQLAFYS ODWSaiffi fcSAftKBSGGS ÍÂKYHMASE QLKRLGQSUC FVLEDLRQ5L imusiKwe luuutntAsx 240 * A 265 2 19 SEQ ID NO: wamriAv lsitq $ qarh fbqqmbosp wmrnnav γνοΑπαιββκ dyvaofeasa ®O DM ^ unuani LesfÊ 'vmwim ΜΒπαηικ sfm' c®Kg mmieaçv 120 OHL-DBPQM 'QESMEmQ' ^ EL APLGHBIR EGMQSft SÊKLsPMEE IEDSÍjRMVF IFTO M & QWkm $ maQWWAR FEALKEGGGS LREYQRKAQE QIKALGgKAK ΜΙΕΟΟφδΙ 240 LPVLENLKVS ILM1DEA.SK 'KLMQ 26S SEQ ID NO 20: MKAALLTLAV LFLTGSQARli PSQQESPQSP WDSVKDLATV YVMVKDSGR DYVÃF EASA 60 IGKQLHjuKLL BHSÍDSLSSfV TKLREQIGPV TQEESÍMjEK BTEVLBQEMS KDLESVKQKV 120 QPYLBOFQKK KV &amp; PÍ.GS &amp; LR SGMftQg &amp; QSL · ÇBHLSPLAEB LRBRftm ¥ B 180 ALR &amp; QLAPYS DDLREHUAR ÁSALKEGGGA SLMYHRMS ESLSM &amp; SKA RPAL3BI &amp;. HQ 240 Í.LPVLESFIW SUMIDEM KKLMAQ 266 SEQ ID NO 21: HKAWX.TMV LFLTCSQAM FWQR3EFRSS ÍTOXÍKBf &amp; W YYBTYKDSSft EYVAQFEASA 60 FGKOLMliíLL DMD5LSSTV SKLÇSQLSPV TQEFWDNLEK KTEGLREE ^ KDLQEVRQKV 120 GPYLBEPQKK 'QEEVERYAQ KVEPLÇAFLR ESMQKLTSL QEKLSfLMS IROSftaXHVS 180 UPVJUSSFi' SróSVLOEAT KKLÍ4TQ 200 SEQ ID NO 22: HKAmiLAV LFLlGSQAKÍi mQQg &amp; FQSS fDSVKQLANV YVDftVKESGR EYVSQLEAS &amp; 60 XGKOmKXV 01 &amp; JOTL6S7F QKYHmGPV AQFFHFKLSK STEELRRElfí KDLSDWQKr 120 GFPLDK1QKK ftQÊOLERYSQ KVEPLSAS8.R EGAPQKLMSL QBQVTPI &amp; SB mmWkXMi 180 TLRYQLAPYS IXftKKTLGAR LEMKEGGSA SLMY8AKAS EQLSALGEKA KPVLEDIKQG 240 LMHfflffiSFKT OTUÍVIBSM KKXTA 265 SEQ ID NO 23:, 8K 'WLAM VFLTGSQMiH VKQQOEPQSQ' DKVKBFANV YVDÃVKDSGR DYVSQFESSS 60 LGQQLtiLHLL EOTDYLGSTV QQQRXGPL JROFÍÍDULEK ET & PMM KDLESVKQEV 120 QPYLDEFQKK mOEOVELYBQ mPtâftELG ESAFOKlQSl QGRLSEVMEWWMRXSVD 180 ELRTQLAPHS KaMRSStW LMLKSfíPTL HBYflTRMCtfa LJSTtSSKAiRP ALEDLRBSLM 240 PMMTLKT &amp; QSVTDftASST LTAQ 264 SEQ ID NO 24: MKRfcVLKVXL VFLTACQMB SHQQOEPQ5Q WDgVKDíATY YVMTKOSGR DYVSQSKS3T 60 ΜΚ5ΜΙΙβ £ ΟϋΒΒΠβδΤν GRLQSOXGPV TOEFKMÍLEK ΈΖΜΙΜΕΜ KCLSIOTQIW 120 ommmx. wmmsm.® KtEPwms khmekqrhl kwmsfr® METODAm i &amp; 6 KFGLYSDQ EífLâORLTEl MBPTLim ® TK &amp; GDSLRYL GEKAK.?AT,CD LGQG1MPVLE RmKEHSMl OEMKKLM 240 259 25 3 SEQ ID NO:! f OEMSYtfBQI KOHLTVYVD AKDSGKDYLT SLDTSALGQQ LKKKLÃDHWD TVSSALLKAR 60 EÇMKFISMÊF 8GMJEDTEG LRQTVSKDLE LVKEÍWQm DSFQKKVBEE LELYRQEÍVAf 120 LSAEWREQAR QSOOTSOKA GSLGOTRDR V &amp; Wit D1APYGEEAB KLLLQSiQSI ISO KAKSGE &amp; AEY QmsmtíS FGEKAQPTLQ DLMGLEP1W EGIKMAMStf LE &amp; LGKKLSS 240 Q 241 SEQ ID NO 26: mWLVfLàV LFLTGTQARS FKQRDEPQÍJT tíOÃXRBMVDV YLETYKR3G'K DAIÂQFFSSà 60 VGKQLDLKLA D & LBXLSRM MLREDÊÍAPY YKBVSEMWLK DTEALRMLT KDLEEWEKI 120 RE.FlDQFSAK "TEELSQYRQ ΒΕ, ΤΡνΗίϊΚΗί E &amp; TKQfímM O & KLTPVASE ARDRLRGHVE 180 SiRKHLAFYS 0ELP.Q: KÍS0K XAEIREKSXF Qkm &amp; UCW EOLSHLREKJÍ TFOTQEFBER 240 LTMfAfiMUffl Rlimtu mm 264 SEQ ID NO 27: JIP-GVliVTLAV LíXTSfQMS STODDP0TT LDRlRDMLOV YLOTKASSK OMSÕFESÊà 60 VGKQLOtm, ÍMimS &amp; tò AKLBEDKTPY YROTREMWUK DTMLRAELT KDLEEVKEKT 120 RPF LBQFSAK TOEFSGTRQ KÍAnmSM OimãWSM QMLTPVAEE VRQRLSEQYS 180 ELRKN1APYS SmteKLSQK LSSIRERG1P QASEYQMW EQLSSLRIKH XP &amp; VQBFKSa 2:40 LTFYMNLliM Rubu &amp; EYO ΚΙΜΑ 264 SEQ ID NO 28: "RWVWIAL · LFUG2QMY HVQBBBEQtf LDRLR01YDV YlBieVK &amp; SGK M1AQFEASà 60 VGKQtDtíííA DNLDTLGMA J &amp; 8SDMAPY YKEVmMRLK DTESURmX KDt.FSVKHKJ 120 RPPLMf SM WTEEIEQYRQ RiapVJfflSLR ELTKQK ¥ EI 4 QQKITPVAEF MDKgGHVe IflQ ELRKSIAPYS DSLRQKLSQK MCRERGIP QAAEYQAKVV SQLSHLRSKH TPLVQDFKEB 240 LTPmiíIKf RFISLiraSLQ Km · 264 SEQ ID NO 29: tíKFIi &amp; LMíTI liMGTOMÍ ^ ÃDAFSQLÉ HVKRALSMn AQVKLTAQRS II &amp; LKZFEYK 60 EYKKQLFQSL WLQGmo & T SQStAPYSBA KSTQIflWC * AVKNBVtKQV SElASQLEfK 120 RAKtMVLBK HXOmmB nm. RTEMEAmK KEPIvmRA W '3MBETJt 180 m & MVEIV RRKLTERLEE mflMPYM EYKEQMIKAV SSVREKVSPL SESFKGQVGF 240 MEQAKQKLL AFYmSQM KA 262 SEQ ID NO 30: ΗΚΠΛΙΛ &amp; ΪΙ LUMTQAVP ííSRDÃPSQLS HlWAfSSEYM ftQVKSTQQRS It &amp; tíHXfêflít 60 EYKVaLEQSL OWLQQYAÇfTT SQSLAPYSEA FGAQLTDãM XIShZ% mW EDVRTQySPK 120 mzimmm HioeYsmB phmivkc® rtsleafrvk mepyveemra kystweetk ιβο AKLMP1VEX? RàKLTERLEE Υ, ΕΤΙΛΑΡΥΑΕ EYKEQííFKAV GBÍfttEKYGPl * TNDFXGQVGP 240 MSQAKÊKLM DFYEfXSQAM KA 262 4 SEQ ID NO 31: sufutax ϊ.ια '6ϊο &amp; ρρ' sacafSíM evkmmmyi WMMS ϊκαμμέυκ 'the SYKMQLSQSL mÓQFADST SKSWEFÍFM SAESCDÃTAT VMSWESSVE DVETQX.EfKR 120 AEtTEVLNKK IDSYMKLÈP LXKQHISLRR TEMDAFRMI DPWSEMRÍtK V &amp; VWEETKS1 180 KU #! VE1W AKLTERLEEt Κ "λΜ" ΙΒ SKEQMFKftVG SVSEKVAFES EDFK & 4? 4 m m m mBBBBBBBBBB ut ut ut ut ut ut ut ut ut ut ut utMP 255 255 255 255 255 255 255 255 255 255 255 255 255 255 255 255 255 255 255 255 255 255 SEQ ID NO 32: MKFVSIM.TI. LLftLÇSQMiO FQAD &amp; PTQLE SYmMLVYl fíSVKEQAEKA LDMLDGTDYE 60 L * K &amp; Qt, SS $ t TKMBYftSTC SQAIíTPYAET I5TQLMENTK QLRERVMTDV EDLRSKLEPE 120 RMiYmOK 810EYREKLE PVFQEYSALS BQffiEQLRM OEPMJDIRK AFESBIEETK 18 SKWMVEAV RTXLTERLED ΧΟ'ΓΜΜΡΥΑΕ EYKEOLYK & Y SMREKIMH TQDLQTRMEP 240 YMEUmTTFA QMYETIAKM OA 262 SEQ ID NO 33: MKFMtAIAt, It & OGSHSÃS MSftDAPSQLD S & RAOLDm tQVKOMSLM VKQLDDPQYA 60 BPKTLLORI ΕΕΜΥΤΟΙΚΤΪ. OOSVSPMTDS FYSÍTVMEVTK DIRSSLHVm EA1KSSLAPÇ 120 WEQLKQV1EK HLNDYRTLL? ntmtSm DEEMAftLKYR LEPVMELRf KIQASVEETK 180 AVIíMMVETV RfRVTERLES LSEW02WQ mEQMRQSflf DOAQWDTDft LRTKITPLVE 240 mVKMSAIF EIIMSVTKS GCG SEQ ID NO 34: MPIKamTL ALVSmQMA EVSADQVMV MWDYSSQLSH NAKEAFEHLQ KSÊttQQtiíA 60 IF ^ BKLGEW TYAGDIOSSKt VPFATELHES LAKOSEKLKl ÊIGKELEELR ARLLPHAtJEV 120 SOKIGDSIRE IíQQRLEPYAD IQ ^ TQVNTOA KQIRRQLTPY ΜΚΜΕΚΠΑΕ H & D $ LQML &amp; i§0 PHAOEIKRKI DQHVBELSSR IffPY &amp; OEFKV KIDQÍVEELSí RSIAPYAQDT QBKLBHQLSG 240 UF0MMAS EIKAKISASA ffiRQRim MPVRG &amp; MG MTK &amp; QKSIA EKKHL &amp; QQV 300 ERA ^ RSVEPY ÇPMKRtVO '-ÍEQORQKLG PHAGDVEGHL SFLSKDIRDK tmSFFSTFKS 360 KSOTKTISL PELEQOQS00' OOQEQVQM L &amp; PLES 396 SEQ ID NO 35: ' FLKÈWDTL ALVAYTGARA EVSMOV &amp; TV M 'DYFSQLSS SAKEAVBHOO KSEOTQC &amp; M 60 LFQDmSSW mGOtÔKKL VPPATEUíEK IAKDSÊKME EXMELEEVR MLDPHAHEV 120 SmGEWRS xmmwrrts QLSTQWST EC &amp; RRQLTPY &amp; 0 &amp; MSKVE8E SADSLQTSLR ISO FSADOLKAKX D2HV1ELKER LTPYADEFKY KLT &amp; WÈELR RSyvHAODA QE8LBHULEG 240 LAFQ &amp; KME ElKARlSASA SEU3QRLAPL MIPMBGMjRG SfYEGLQSSlft SLGGHLDRHY 300 ESFRI.RVEPY GSNFKKftLVQ QMEQLRQXXG PHAGPVEGOL SFLG &gt; &lt; DLSSK VSSFFSYFKÊ 360 kesä ^ TLSL PBFSQfiEEQG QEQQQSQSQE OQSOÍfiQQQQ QEQQ8EQQQQ EQSQEQQQBG 420 Vq 'L &amp; ple $ 42S 5 SEQ ID NO 36: fcFLKfiAVXTL acvftissxafc SVTSDQVMV VWDYETQLSiSi WmMEQFQ KTDVTQQLST' 60 LFCDKLGCAS TYADGVENKL VPPVVQftSea UU &amp; TERVKE EIKKSLSDLR BSMMFHMKV ' 120 TQTF6EBM0K LQEEMFYÂV DLQDQXFÍQT QMKL01 * TPY IQMQMKE BVIMKRSHT 180 FIAmKDISP DMM LT & FÃMMLKFT TIDQSLEDlift SSIAWaVS? QEKX23 £ Q $ g £ G 240 I *? CBKÍK * I »BE®WS.ffiI W2LQKÍÍLML VECWDSKVKG FTSOLOKSLE BLSRtaEQOV 300 EEfRKmp» SSK8WRI.VO «^ 5 BISGEVBSHL SM45KSIREK VHSfMSTLM 3« 0 KGSPDQFOftL PLPEQAQEOA GSQ & QEQVQP KPLES 3 95 SEQ ID NO 37: CMÃEvsftOQ vmvhkmfs QhBèwmm ehlqkseltq qluMjFqdkl gevntyagbl 60 QKKLVFFATE LfiERlAKOSK KLKÊEIRKBL EEVRÃHLLPH MEVSQKIGE KVmfiQRLE 120 PYTDQLRTQV NTQTSQL8S.Q LTPYAQRMRH VIREHADSLQ TS1S2SADQL KAKIDQ &amp; Vee 180 LKSR1TPYAD EFKVKIDQTV EELRRSLAPY AQffiQÊKLHH GtSPtAStíMK KSAmKARI 240 SASAEELRQS LAPIAEDHRG RLRGOTEGtQ K5LAELGSKL DRHVEEFRLR VEPYGSNFNK 300 MffOQHBffi * QKLGPKAGW EGHL5FLEKD tROKWSFFS TFKSKESODJSÍ fLSLKFBQÔ 360 QEQQOTG2 QEGQSEÔGQÔ SOQQEQEQQQ EQVÇMAPLS $ 401 SEQ ID NO 38: HFLKAWLSL ALYAVTGMA EWADQmTV MSDYFSQLGS MKK &amp; YEHLQ ISEtTOSIiST 60 tlODKMKVN TYIESLífKKL VPFATSLME tTKOSKKtSE 8ÍRMtE £ 120 liR ASLEFRATEV 3QKIWNVRS WXmGFFTQ SLRTQTOTQV QQLQRQLKPt AERMJESVLRQ NIRNLRASVft mOÊEKAÍO 180: BQNVEELKSS LTPYMSiKA íílDQfíVEiliR RSLAPYAQDV Q mMQLEG 240 IMgtiMSW BWMISMA OELROKIVPV AEHVKGIS.SG 'EtGMRSÚ, IIJtôaiDÔeV 300 EEFRLKVSFY GETFHKftlVQ QVEOLRQKLG PIAPDVFGftl SFLSKDLRDK WTFFSTLKE 360 EASQCQSQAL PAQBK &amp; QftPL SS 382 Θ SEQ ID NO 39: MFLKAVVLiV ftLVAnGTQA EVTSOQVMV HSHYFÍQLSÍi NAKEAVFJ &amp; Q KT0VTQQL8T 60 tFQDKLGSXN 1TADOLQÍIK1 VPPAVOLSGH LTKSTERFRE EIOKELEDLR AHMMPRARKV 120 SQHFSDSVOK LSBaiiRjPYAT B &amp; QAQINSQT QMKRQLTH IQHMQTTIÔD HVmQSSHV ISO PFMELKEKF SgKMEGUNGQ tTPMÍJEÍLKK T1DQÍÍLEDLR SRlAPtMGV OSmiSCMÍG 240 IAFQJ 'K8AE ELGWSTM Wtfmiit &amp; h VEDYQSKLEO ETESLQKSLE DLMQtDfóJV 300 IWBM ^ SPL GOKílMALVÔ QMMFRQQLG SDSGDVKÊ8L SFLEKE1.RM VSSFH5TIQK .360 KÈSPDQPIAL PLPEQVQEQV Q2QVQPKPLE £ 391, and (i) X is a protein heterologous heterocycle comprising more than 200 amino acids, provided that in the case of the construct it consists of two identical apolipoproteins exactly 6 the same, then these are attached from the C-terminus to the N-terminus, or (ii) X is a heterologous radical comprising a the oligomerization, wherein the construct has an increased plasma half-life compared to the wild-type apolipoprotein; and wherein the analog or variant is capable of eliciting the same physiological response as apolipoprotein-A-I, A-II or A-IV. A construction according to claim 1, further comprising a spacer between the apo-A and X component. A construction as claimed in claim 2, wherein the spacer comprises a peptide spacer. A construct according to claim 3, wherein the peptide spacer comprises at least two amino acids, such as at least three amino acids, for example at least five amino acids, such as at least ten amino acids, for example at least 15 amino acids, such as as at least 20 amino acids, for example at least 30 amino acids, such as at least 40 amino acids, for example at least 50 amino acids, such as at least 60 amino acids, for example at least 70 amino acids, such as at least 80 amino acids, such as at least 90 amino acids, such as about 100 amino acids. 7 The construct according to claim 2, wherein the spacer is connected to the apo-A component and to X through covalent bonds. The construct of claim 2, wherein the spacer is essentially non-immunogenic and / or is not prone to proteolytic cleavage and / or comprises no cysteine residues. A construction as claimed in claim 2, wherein the three-dimensional structure of the spacer is linear or substantially linear. The construct according to claim 2, wherein the peptide spacer comprises the tetranectin GTKVHMK amino acid sequence, the human fibronectin binding chain PGTSGQQPSVGQQ and GTSGQ amino acid sequence of the murine IgG3 upper hinge region PKPSTPPGSS , SGGTSGSTSGTGST, AGSSTGSSTGPGSTT or GGSGGAP. The construct of claim 1, wherein the protein or peptide comprises at least one mammalian protein. A construct according to claim 9, wherein the mammalian protein is a human protein. The construct according to claim 9, wherein the protein comprises at least one protein selected from the group consisting of albumin, more preferably serum albumin, the plasminogen serine protease fragment or other serine protease manipulated so as to be inactive by disruption of the catalytic triad, and the immunoglobulin heavy chain constant region. A composition according to claim 1, wherein component X comprises at least one apolipoprotein AI, apolipoprotein A-II, apolipoprotein A-IV, apolipoprotein E, an analog or variant thereof which shares at least 70% sequence identity with a sequence selected from the group consisting of SEQ ID NO 15: 60 ion ISO 240 267 SW 120 180 240 267 60 120 ISO 240 2E7 mmnfim lfiksq &amp; k fwqqdeppqs pkdrvkdlm vyvbvusdsg rejyvsqfess lítMOSVtSÍ fSKLRECLGÍ VTOEÍlfOTLE KETEGLRQKÍ4 SKDLEEVKAK TOHLMWQK KWQ8EMBLYR CKSffifLRm QfiSftBQlCUig lgeklseloe emsdmísmv SDEMOSLM RLERLKEMSG ARLAE'YHAEA TSHLSTLSSK MPAIEBÍ 'GhLPVLESFK VSEWK &amp; LKBY fSKÍÍÍTO SEQ ID NO 16: HKR &amp; VLTLM LFLTSSQARH FWQQDEPPQS PWDRVXDMT VYVtMiKDSS EDWSQFIGS aL®®PfI8St LDWTOSVTST F5KL1EQ1GP WQBSWHBJi KETEGLRQSií SKCLBEVK & K VOmDDFQK ΜβΕΕΜΙΪΚ &amp; ΝΖ' υΒΜΙ QBGÃSÔKLiíE LQSKLSPÍ6É EM3DRARMV mmmiMH δΟΕΙβΟβΙ &amp; Α RLEALKEKGG ΑΚΕΛΕΑΚ Κλ TEBltSTLSEK JUffiALEDtSQ SW.PTOESFK vmSMiEM SEQ ID mLltTQ NO 17: MKAm LAV traGSOSBH FWQQDEFPQT FK3RVK0 &amp; VT VYVESLKDEG KDYVSQFEGS MJSmXX * LDSÍWB3TOT V3t &amp; REQtiG * VTQBmWLB KBTSSbRQIH SKDLEEVKAK vomo »ITAC qkveplmèl bestrqkiie lhekxsplge e71orarahv E &amp; LST1LAPY SOStRQRIM SRLAEYHfiKft SEHLSTLSEK MPALEDLRQ G &amp; LPVLESFK VSrtSA.t.tEY TK &amp; LSTQ 9 SEQ ID NO 18: MK &amp; YVLTLAV trLTGSQSRa fSSQC &amp; gQSS WDSVKOFATV YYEMKDSGR BWAQFEASA 63 LG & QLNLKLL DMDflASTL SKVREQLGFV TGEFMHftSX KDLBSVKOKV 12G QFODBfQKR '®S35V8X' RQ J '&amp; ífcGESFR B ^ QRW. ODKLSmQS UUftHiAWK ISO TLBQQLAPYS SMQRLTM LEALKEGGGS LAEYHRKÃSE QLXALGEKAK pyiEOLRQGL · 240 J WJR $ HWS TtMJPSftSK KLMA &lt; 2 265 19 SEQ ID NO: MATOTIÂV LFITGSOAM ÍWSfDBPQSP 'OBVK' RATF WtmiKDâGR DYVAOFSRSft LGMMLRLL 0MÍ5SÍGSYÊ 60 'fSWMQLOP? tQÉFSDSIIlK EXEALHQEMS KBLBBVKKKV 120 OmDOFQNK WQSEMETYRQ KMAPltGftEFR EGAR ^ / QGL QÉKL $ FÍAEE LROMcRAHW ISO ÃLRQtíVAPYS DOlSmMMR SmKEGGGS LASYQAKAQE gLKALGEKAK FALEDLRQGt 240 LFVX.EMLKVS IL &amp; MOEASK KLSfAQ 265 SEQ ID NO 20: MK &amp; ALLTIAV LFIíTGSQMR BvQCPSPGSP WpRVKDL &amp; TV YVD &amp; YKOSGR BYVAQFEASft 60 IGKQLNLKLL · ^ ISK DKfíOSLSSTV TKLRSQKSPV TQEFWD ETEVLRQSMS KDLESVKQKV 120 OPYLDDFQKK WQSEVEmO KVÁFLG3ELR ESARâKLGSL QEKLSPLÃEE LRD8AKT8VD 1oo M.MQLAPYS DS | jR.EKDMR ÁSALKEGGGA SLAEYHARftS EQLSAL5SKA RPALEOÍ HQ ^ 240 LLPVLSSFW SXiRÃIOEÃT KKLNAQ 266 SEQ ID NO 21: WIAM / rLftV & ÍLfGSOflAH FDQRBEPRSS WDKXRBF &amp; F ? YVBTVKD3GR EJVAQPEASA 60 FGEQLMLSLL WÍWD * $ SSW SKLSBQ5WPV tQEFWQNLEK ETEGLREEÍSS KBLQEVRQKtf 120 QPYLDEFQKX WQKBVERYRQ KVEPLGKELFl ESARQKLTSL QBK &amp; J. SFJAES LRPSMHV3 180 FVLSSFm SVQRVLOEM KKLOTQ 206 SEQ ID NO 22: wamiuw lflygscarb vtmmmss idrvrdlakv yvejwhessr ewsglmsr. are LGKQliíIKLV DMWBTLG3TF GKmmGPV AQSFWEKLEK ETEELP &amp; 2IM KE &amp; EDVRQKT 120 QÍFIíDEIQKK SQEDLERYRO KVEELSAÔtR ÊGMQBIMEL QeQVTPIGSD LRDSVRMAD ISO TLRTCLRPYS SOSOCIZSMt LMÍKEGGSA S1ASYHSKAS EQLSALGSKA KTCUCXHQÇ 240 I.MPM8SSFKT SVLHVIBBAA KK1TA 265 SEQ ID NO 23: WaWtAVJUl VfLTGSQA8K VBOQDEPGSQ SDKVKDIANV YVDAVKDSSR DYYSQfMSS 60 EGQSLKUIU EMOTLGSTV SQLQSRLSPt TROPKDNLEK ETDWVRQEí® KDLEÊVKQKV 120 QPYLBSFQKK KKKIWELYRQ KVAPLGAELQ BSARQKLQEL OGRLSPVME FRDFJ3RTHVD ISO SLRTQÍAPBS EQHRESLAQR IMUSSPTL ΝΕΥΜ'ΜΚΪΒ imGSKARF AimMSlM 240 PMLETLKTKR QSIAIOSASKI LtAG 264 10 SEQ ID NO 24: MhWUW &amp; l VFLTéCQMÍE S ^ QQD.EPOSQ »DS ¥ K3 &gt; mV WMVKCSSR DYVSQFESST 60 ISMmJSU, DHBBTLGSTV GKLQEOtXSPV TOEFKAIÍLEK ETDSERNEHN KOLBMWQKM 120 QPHLDEPQEK WHEEVEAYRQ KlSíWmB KRAKEMORBL KWRgEFRPR KRVSADftLR * ISO KFGLYSDQMR ENMQR1TEI RBHPTLISYÍí TK &amp; CDRLRTIj SEKAKEALDD LGQGLMFVIE maaama bbbxxxim 240 259 25 SEQ ID NO: OaaKSYÍTOQI KDMLTVYVM 'AKDSGKOYLT SLDTSMiGOQ iMMD TVSSALL KAR 60 HS 'ffl®ISMEF WaffiUBKDTEG LRQSVSKDLfc LVKEKVQPYL DSFQKKVEEÊ LS ^ XST & m' 120 LSAEWRÊQAR 'XAQFXÇQKft GELGQQHRDR VRTRVDALRT DLRPYGEERB KLLLQRLQDI 180 KAKSGEOEY QTKLSESIKS FGEKAQFTLQ RLRgÇLEFLW EGIKSÍSAMSW LE &amp; LGKKLKS 240 Q 241 SEQ ID NO 26: MRQVLVmV LFLTGTQMS FKQHDEpgrF LORIRMVOV YLET ¥ KASGX DAIAQFESSA 60 VGKQLDLKLA DlíLsTLSAM MLREImj Y YKBVREÍWLK DTEALR &amp; SSS SDLEEVKEEI 120 RPFLDQHSM OTEELB0YRQ &amp; LTFVAQELK ELTKQfW £ Ii &lt; 8 QMLTPVASS ISO MDRLRGHYE SLRKKLRgYS OSLKQKLSQK LESIREKGXF QP &amp; MQMM SQISHLREKH TFLVQEFRER 240 ItTRYAEKIíM RLISFLDELQ KSVA 264 SEQ ID NO 27: MR6VLVT1AV LSXXGTQMS WÍKDtJWf LDBIRDMWW YldSCTKRSSK DAISOFBSS &amp; 60 VGK0LDLKLA Oí ^ mSAAA AKLRSBKTPY YREVftMLK OfEALHAE.LT KBLEE ¥ Í®KI 126 RPFL.DQFSAK WTEEVEQYRQ IIAPTOSEM PLT &amp; QKVELM QRKLIPVME VKmSEQVS 180 EI £ XNXAPY8 SELRQKLSQK LESIRERGIP QASEYQAKW EgiSSLBEKM TPLVQEFKEE 240 ITPYMÍÍLKJ &lt; I tut &amp; imn KZM 264 SEQ ID NO 28: MRWWFLAL LFLTGTQmi FMQBBBEgtt LDRLRJJLVDV Ytt & fVX & SGK BAIAQFEASA g &amp; VGKOLDLKLA DNIDTLGMA AKLREMAPY YKEVREMBM DTESLRAELT KDLEEVKEKI 120 MiIMUM, WIEELEQYRQ RLAPVAEELEÍ ELT5ÍQK2ELM Q0SMPVAES! AKDRLRGSW 10Q ELRKHLftPYS DSLRGKLSOK LEEXM3KGIF QAAEYQAKVV KQLSSLRSKM TPLVQDFKER 240 LTPYMTLLKT RFXSLIDSLQ KT? 264 SEQ ID NO 29: MKFULALTI LLMGTÔAFP MQADAPSQLÊ BVKRALSMYI AQVKIfAQRS XBLLODTEYK §0 EYKMQLTQSL DNLQQYADAT EÔSLÃPYSEA FGTQLTDftTA AVUAEVtKDV SELRSQLSPK 129 MELKEVLDK HIDCYKKKUS PLIKEKIELâ RTEMSAFRAK MEFIVSELRA KWUCIWBBIK 180 mAÍPIVSIV RMLTgRlSE LRTLMPYM EYKECH1KAV GEVREKVSPL SEDFKGQVGP 240 262 KR MEQMQKLL AFYETISQM 11 30 SEQ ID NO: KKFLÃLALTI LLASATQAVP MQftDAPSQLB HVWAÍWYM AQVKETGQRS mLSmflM 60 EYÍWQLSQSL &amp; NLQ3YAQTT SQSimSEA SSAT &amp; TDAAA fflMSVMKDY EOTRYOÍ.ÊPK 120 SMLKBVLDK HIDEYMfOtE ÍÍUMIVEQR RmSftPRVS 'BPWEE "BA K7STMVEBTK 180 JWOmvm RAKLTESUEE LRtXAÃPYAE EYKEQ8FKAV GEVRSKvm TifDFKQQVGP 240 .imQkXMLX DFYETISQAfí KA 262 SEQ ID NO 31: HKFmftm HAAGTQAFF' QACAPSQIiE SVXMLNHKI ftfiVKLTSQRS SDILDSIEYK 60 SYMQLSQSL ΟίϊΙίΟΰΕΑΕβΤ SKSSmPSS SAPSCOATAT VmEVHKDVE DVRTfôLEPKE 120 mTSVLNKH IDMRKKLBI? 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SFLEKDLPHQ VNSFFSYFK 360 KSSQDMSL PEYEQOQQQ QQQQQQQQQ LQPHQ 398 SEQ ID NO 35: 12 anjawm It is important to note that in this case, ! £ R * wpâm.si lmoseks eiskelsêys, arliíhahbv 120 SQKIGEWRE I ££ ma $ SJ QLRTQWTQT EQLRRQLTPY &amp;! QMSRVM HADSLQTSiR ISO PMÍJOIMKI DQNVEELKER GTPYAB &amp; IW KIOQSmLR RSJÃSXAQBA QÊKLBHQL.EG UQ LAFQÍÍKKMAE ElJáRlSAS 'ESXSQBIAFL KgOtetCKLRS WTXGUKSIA SLGGHLDRHV 300 EEFELRVEPY GSHFKKMYQ C ( M £ QLRQKt &lt; 3 PHÃGOVESBÍ SFLgKOlSSR VBSÍfStm 360 KZSQmUlSl, PSF6Q0REQQ QEQÍiQEQEQE OOOQQEOQÚQ QEQQ8EQQQQ EQQQIQQQES 430 V ^ ORS 42Q SEQ ID NO. 36: ^ FLKAAYLTL ACVAITGT8A ÊVTSDQVMV VStDYFTQLSN SAXEAVEQfQ KTDVTQQLST 60 ItFQOKLGOAS lYADGVHNKL YFFWffiSGH LMETERVKE ElKKgLHJLR DRMMFHMKV χ20 TQTFGEUtíQK l0SOKfíkV DLQDQINTQf 0S £ M, Q1 &gt; TP ¥ ia "fission OTmOTâM ifl (j PLATNLKCKF 8SSKEEIKGH LTPSANELKA TIOQfSLÊOtt SSMIPMVGW βΕΚΏΜΟΚΕβ 240 IAFQ8KKS 'and ELQTKYSMI DflLQMÍIML TOW3SKVK8 HTSGlâKSLK 0' 0:! Lg &amp; I V 300 EEfRRmPS SSMP8IOLV0 QLEOS ^ QQtS WÍSGEVBSHL SFLBKSIMK VHSFSJSTMK 360 RGSPDQPQAL PLPEMEQA QSQMSBQV0P KPLSS 395 SEQ ID NO 37: GAMEVSRDQ VATVMKOIRS QÍSSSSMKEM mÇSEMQ Q LMLFQDKL GSVSTYAGBL gQ QKKLYFFftTB LHERLÃKDSK KLKEEXRKB1 REVRÃHLLPH ÃQSQKXD. KVSSLQQROI IGQ P1TDQLOT? OTQTEQÍBáRQ LimORHES VLRHKA &amp; SLG TSIÃFSAÍJQL · KAKXDQSSVEB IGQ MWMmftD ElWfKlfiOTV EELBRSIAPT AQMQEKIKH QUEGUUTGHK fôíMELKARI 240 SASAESLRQS LftBIMDHRC m & SHTES &amp; fl KSMELGGHL OREVEEÊRIR VmGEMRK 300 ALVQQ ^ BLR QKLGPMGOV EGHLSFLEKD mmm &amp; TFB tFKEK &amp; AQI 'TLSLPEPEQQ 360 QEOQQESEOQ QBQQEEQOQO SGQOEGEQQQ EQYÇMAFLE S ¢ 0.1 SEQ ID NO 38: MFLKÃWLSIi ALVÃWSÃM EV8 &amp; CVD &amp; TV &amp; TOYFSQLS5 WPEHLO KSELTOQtaT GQ IFQtiKbGSVK TYTEGLQKKL · VPFÂTELHEE 0TKDSEK1.RE SIMELÊÊLR MLWHffiEV 120 SQKISD8VRS LQQRLGPFFG GLftTQVMTQV QQÍ ^ RQI-KRY AERMESVLRQ NiRtíLE &amp; $ VFT χ§0 mosassati dqkveelkgs ltpymbIíKa kidqmvssea rslapyrqbv QEK &amp; nhqleg 240 mmHg mmHaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaa SFLEKDLROK YMSFFSTLKE 360 saaososoai. mMwmm m 3δ2 and SEQ ID NO 39: KFLMWIW AMMIGFQS. EVTSDQVMJV HHDSrFTaWW MKEAVK &amp; Q KEOTTQQLHT IFQDK6G8IM TYADDIQÍÍKX 60, VPFAVOISGH LTKSTERVRE EIQKEIEDLS, 120 M1IHH1À8KV SQKFeDSVtiK ΙβΕΪΒ, ΚΡΥΛΤ Μ0ΑβΙΜ '0Τ δΟΚΚΚ &amp; ΪΡΪ IQRMQtfXQD HVBHLQSSMV PFMELBEKF 180' gNHEÍ &amp; KGO X / fPRMELKA TIDQ8LE0LP. SRSAPIAESV OEKLÍJOQMEG 240 ΙΑ, ΡΟΜΚΚΚΑΕ ELQTXVOTI DÇ & QM & PO VELS ^ StaáKS STESIiQKSLE SOSKQLDQQY 300 IWBMYSPL GDKFSmtVa QIBSFRQQIJG S05SB «rgS8I» SHFâQKI®K VSSIBSTW »360 KGSPDOPLAL FLPEQVQEOY Q20WKPIE $ 391 13 A construct according to claim 9, wherein the peptide comprising component X comprises more than 300 amino acids, for example more than 400 amino acids, such as greater than 500 amino acids, for example more than 600 amino acids, such as more than 700 amino acids, for example more than 800 amino acids, such as greater than 900 amino acids, for example more than 1000, 1250, 1500, 2000 or 2500 amino acids. A construction as claimed in claim 1, wherein the oligomerization module is a dimerization module. A construction as claimed in claim 1, wherein the oligomerization module is a trimerization module. A construction as claimed in claim 1, wherein the oligomerization module is a tetramerisation modulus. A construction according to claim 1, wherein the oligomerization module is non-peptidic in nature. The construct of claim 15, wherein the trimerization module comprises an amino acid sequence, which is capable of mediating recognition, trimerization and alignment between the three polypeptide chains. The construct of claim 15, wherein the trimerization module comprises the trimerization modulus of tetranectin obtained from tetranectin. 14 The construct of claim 19, wherein the trimerization module of tetranectin is capable of forming a stable complex with other trimerization modules of tetranectin. A construct according to claims 19 to 21, wherein the trimerization module comprises two trimerization modules of tetranectin bound by a spacer group, which allows both trimerization modules of tetranectin to form part of a complex with a third trimerization module of tetranectin that is not part of the apolipoprotein construct. A construct according to claims 19 to 21, wherein at least one of the trimerization modules of tetranectin is selected from the group consisting of human tetranectin, murine tetranectin or human cartilage, bovine or shark C-type lectin . A construct according to claims 19 to 22, wherein the trimerization module of tetranectin comprises a sequence having at least 68% identity to the consensus sequence of SEQ ID NO 12. The construct of claim 23, wherein the cysteine residue # 50 is replaced by a serine residue, a threonine residue or a methionine residue. 15 The construct of claim 23, wherein the cysteine residue # 50 is replaced by any other amino acid residue. The construct according to claim 1 which has a half-life of at least equal to the native Apo AI, A-II or A-IV, preferably at least 2-fold, more preferably at least 3-fold, such as 4-fold, more preferably at least 5-fold, such as 6-fold, more preferably at least 8-fold, such as at least 10-fold. The construct of claim 1, which has a higher cholesterol binding affinity compared to native Apo A-1, A-II or A-IV. The construct of claim 1, which is capable of binding to a receptor selected from the group consisting of cubilin, &quot; scavenger &quot; Type 1, Class B (SR-BI), &quot; ATP-binding cassette 1 &quot; (ABC1), lecithin: cholesterol acyltransferase (LCAT), cholesterol ester transfer protein (CETP), phospholipid transfer protein (PLTP). A construct according to claim 1, having an amino acid sequence that shares at least 70% sequence identity with one of the sequences SEQ ID NO 3 to SEQ ID NO 11 or SEQ ID NO 14. Use of a medicament as defined in claims 1 to 29 for the preparation of a pharmaceutical composition. Use according to claim 30, for the treatment and / or prevention of atherosclerosis. Use according to claim 30, for the removal of endotoxins. Use according to claim 30, for the treatment of angina pectoris. Use according to claim 30, for the treatment of myocardial infarction. Use according to claim 30, for the treatment of angina pectoris caused by atheromatous plaque. Use according to claim 30, for the treatment of unstable angina pectoris. Use according to claim 30, for the treatment of arterial stenoses, such as claudication, carotid stenosis or cerebral arterial stenosis. Lisbon, 12/15/2010 1/23 FIGURE 1 SQ SEQUENCB 267 AA; 3077SMW; 1A2SB83á6E5203I CROCO4; SBQ ID KO.MS MKAAVLTLAV LFLTGSQARH FWQQDEPPQS PWDRVKDLAT VYVDVLKDSG sbYVSQMGS MXjKQIMJCL HJNW0SVTST FSKLREQIX5P VTQEFWDNLE KETEGLRQEJví SKDLEEVKAK vqpylddfqk kwqbemelyr qkveplrael qegarqklhe lqbklsplge emrdrarahv DAIATHLAPy sdelrqrlaa. rlealkengg arlaeybáka hhlstlseb: AKPALEDERQ βΟ ^ νΐ, ΕδΙΚ vsflsaleey tkkiotq 8® «3 &amp; Mee * q «e Bsvtse m Pag Jttftfeit end« h * e «w Ws« K! Bftfc Exit. SsàgeSsc amp sp sp sp sp sp q q B B B B B B B B B B B Br Br Br Br Br Br Br Br Br Br Br. ealR08 tetofifih eea breaa nu ttec ee bwceei3 ee88, ufcfeit 8 'Maus' Kât Bwr. Hedgehog, Mekenen O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O Atl Atl Atl Atl Atl Atl Atl Atl Atl Atl Atl Atl Atl Atl Atl Atl Atl Atl Atl Atl Aalisen Asbysiab Ssa Braam ÍFW5® Maçará © Swiae m oeg Rabblt ttm sfese * MOttes «at £ ur. Heijebeij Cftieken 3ap. FIG. 2a. FIGURE 2a. FIGURE 2a. FIGURE 2a. FIGURE 2a. FIGURE 2a. FIGURE 2a. FIGURE 2a. FIGURE 2a. FIGURE 2a. FIGURE 2a FIGURE 2a.1 FIGURE 2a FIGS. (M + H) + (M + H) + M + (M + H) + M + ------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------- The invention relates to a method for the preparation of a compound of formula (I): wherein R 1 is as defined in formula (I) and R 1 is as defined in formula (I). ϋ α ι Αζ ϊ ϊ ϊ ϊ ϊ ϊ ϊ ϊ ϊ ϊ ϊ ϊ ϊ ϊ ϊ ϊ ϊ ϊ ϊ ϊ ϊ ϊ ϊ ϊ ϊ ϊ ϊ ϊ ϊ ϊ ϊ ϊ ϊ ϊ ϊ ϊ ϊ ϊ ϊ ϊ ϊ ϊ ϊ ϊ ϊ ϊ ϊ ϊ ϊ ϊ ϊ ϊ ϊ ϊ ϊ ϊ ϊ ϊ ϊ ϊ ϊ ϊ ϊ ϊ £ £ £ £ £ The invention relates to a process for the preparation of a compound of the formula (I): wherein R 1 is as defined in claim 1 in which R 1 is as defined in claim 1, wherein R 1 is as defined in formula (I). ATOKYYYYYYYY ^ &Amp; AMISÍSÍJWAPfTrSEV ^ SWIiKDTÇSÍiSAEinSEtJiSBVÍBK ΒΪ- SSfKHQk ^ Mm ^ ftllATSaSU ^ s £ ~ j®w ajgs ^^ b ^ s ^ EI-SAMB 'SMS &amp;! 8 * f ^ S * BSf8 ^ S &gt; ^ dy- KJYKLQlSMttK ^ JStAíjnSQAfcfPÍAKTJSTQlítaSTKJJiMKWfBVEOLRSX ^ -ABVKmASKIBSimÇlK ^^. . . , VOPniffiFQÍ ^ EE ^ YSQKWIiRMtjÕãÔftRQKU ^ MspI ^ maBft & fttlHV VQmmXMQI &amp; Kttnt ^^ VQPYy5PQKKWHgEVBÍYBSK5fA íí6BSFÃÊ &lt; 3AlíQKlf5 $ IÍPÍl &lt; SPliAQEIÍCRÈE®V VQP '^ lí ^ aaEMTm339WQMITOSkRÔKy ^ I ^ M} ^ ft ^^ m' ^ VOmBÍ ^ SW! ^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^ " ^^^^ Α "" ί8 &lt; Ι ^ ^^ ^^ ΚήδΕ νΚΪΗΒΓ ispmiqpsraBimsqsi ΙΒΡΜίΙ ^ ^^ ^ ΑΜίΠίΙ ϊΒ IJSPi * ^ ^^ ^ MliKPWiíkHIEEVRKK iXSffiEIEIátííJBSlEAFSMÍÍKPIVERÍáWlKVAIÍÍV M '' 8E i3ijtiimimm.im ££ ^ ^^ ^ ¥ S8 FKSmrmxHmMmRfc The invention relates to a process for the preparation of a compound of the formula: ## STR2 ## in which R @ 1 is as defined in formula (I). . 2 H); I + * &gt;: f &gt; * mssfis ΚΜβφίβ ewia * Fig. 1 Fig. (a) (b) (b) (b) (b) (b) (b) (b) (b) and (b). The composition of the present invention is described in US Pat. salmo® febrafisí »Sea bream 3/23 FIGURE 2a.2 SSO! KMJim ^ S® | The invention relates to a process for the preparation of a compound of the formula: ## STR1 ## in which the compound of the formula ## STR4 ## is selected from the group consisting of: ImsgSfOKB ^ II ^ KBSGSMLM ^ eyKTíj ^^^ ^ í $ 2 w mpAMSiGíj SEliUÍSiA ÊliPeKI ^^ ^ Q ^ ^ 3SIJQSÈÊÈÍftESáíP BKiRKRIMySSS1 SRrRKroiFQftSKirítWMSQliSSLaESfKrHiVQEKE 26 ^ RS ^ 'ÕAKWB3SiSI (njtSKS5THi ¥ t5KPKS 3 B8] u? U (mHMi8Í QH ^ ^ ^ Sl ^ Ρ fâi®Í ΑΛ ΑΧ ΑΧ ΑΧ ΑΧ ΑΧ ΑΧ ΑΧ ΑΧ ΑΧ ΑΧ ΑΧ ΑΧ ΑΧ ϊΟΡΚΚ ϊΟΡΚΚ ϊΟΡΚΚ ΑΧ ΑΧ ϊΟΡΚΚ ϊΟΡΚΚ ϊΟΡΚΚ ϊΟΡΚΚ ϊΟΡΚΚ ϊΟΡΚΚ ϊΟΡΚΚ ΑΧ ΑΧ ΑΧ ΑΧ ΑΧ ΑΧ ΑΧ ΑΧ ΑΧ ΑΧ ΑΧ ΑΧ ΑΧ ΑΧ ΑΧ ΑΧ ΑΧ ΑΧ ΑΧ ΑΧ ΑΧ ΑΧ ΑΧ SV ΑΧ SV SV SV (1), (2) and (3) and (3) and (4) and (4) and (4) and (4) are as defined in claim 1, The title compound was prepared from the title compound as a white solid, mp 218-221øC. 1 H NMR (CDCl 3):? (s), and (c) (b) (c) (b) (b) (b) (b) FIG. 2b shows a cross-sectional view of a cross-sectional view of a cross-sectional view of a cross-sectional view of a cross-sectional view of a cross-sectional view of a cross- ® »ASM Λ5Μ '' ΑΓΡΙΟ .APMJlAT S3Q m ΜΟϊ HPtKAW &amp; ^^ TMLVAVABA 66 U 35 is saRtómmviaTGmsvr ftijvv ^ ^ fi ^ i ^ ^ v ^ Lo KrDVTQQí.sT: &amp; m &lt; / RTI &gt; &lt; / RTI &gt; &lt; / RTI &gt; (s), 2 (S), 4 (S), 6 (S) Eur-lex.europa.eu eur-lex.europa.eu The invention relates to a method for the preparation of a compound of the formula: ## STR1 ## wherein R 1 is as defined in claim 1, wherein R 1 is as defined in claim 1 in which R 1 is selected from the group consisting of: 8LMYYYYYYYYYYYYYYYYYYYYYYYYYYYYYYYYYYYYYYYYYYYYYYYYYYYYYYYYYYYYYYYYYYYYYYYYYYYYYYYYYYYYYYYYYYYYYYYYYYYYYYYYYYYYYYYYYYYYYYYIYYYYYYYYYYYYYYYYYYYYYYYYYYYYYYYYYYYYYYYYYYYYYYYYYYYYYYYYYYYYYYYYYYYYYYYYYYYYYYYH In addition, the invention relates to a process for the preparation of a compound of the formula: ## STR2 ## In the following examples, the term &lt; RTI ID = 0.0 &gt; Mf &lt; / RTI &gt; ? 27 | ASM BUWSI? | P3ISI? A? KACPA? 9 | 96572 | | α | SpteS758 | A? M_P? | M Bpt 046409? A4? PIS epSP5? 651 M? G? M? 53? 3? C? EXAMPLE 1 S6-C6-C6-C6-C6-C6-C4-C6-C6-C6-C6-C6-C * ϊ ϊ ϊ ϊ ϊ ϊ ϊ ϊ ϊ ϊ ϊ ϊ ϊ ϊ ϊ ϊ ϊ ϊ ϊ ϊ ϊ ϊ ϊ ϊ ϊ ϊ ϊ ϊ ϊ ϊ ϊ ϊ ϊ ϊ ϊ ϊ ϊ ϊ ϊ ϊ ϊ ϊ ϊ ϊ ϊ ϊ ϊ ϊ ϊ ϊ 968728 ASM StmM A9A4je 'K & ASM MOSISE ASM 9IS asmIm pMAt &amp; MMDQNV8' The invention relates to a process for the preparation of a compound of the formula: ## STR2 ## In the following examples, the compounds of the invention may be prepared by the following methods: ## STR1 ## ## STR8 ## ## STR3 ## ## STR3 ## ## STR4 ## ## STR3 ## EXAMPLE 286 FPSS SEQUENCE CHARACTERISTICS OF THE INVENTION The invention relates to a process for the preparation of a compound of the formula: ## STR1 ## in which R 1 is as defined for formula (I) (1) (1) (2) (2) (2) (2) (2) 088409 i952851 ASMMMMMMMMMMM ASM. MPMâ,,Aâ,,Mâ, ..., â € ‡ â € ‡ â € ‡ â € ‡ â € ‡ â € ‡ â € ‡ â € ‡ â € ‡ â € ‡ â € ‡ â € ‡ â € ‡ â € ‡ â € ‡ â € ‡ â € ‡ â € ‡ mm) MMS [mm] ASM mm 1933821! ASM KACSA josiso] asm mms j3281S5ÍA9M "ΡΛΪΜ] 046; 4O9 [M & pie pie 90 90 90 90 90 90 90 90 90 t t t t t t t t t t t t t t t t t t t t t t t t t The invention relates to a process for the preparation of a compound of the formula: ## STR4 ## (ISO: 369); (i.e., in the presence of a compound of the formula: ## STR2 ##) The invention relates to a process for the preparation of a compound of the formula: ## STR1 ## in which the compound of the formula ## STR2 ## wherein R 1 is as defined in formula (I) ## STR1 ## wherein R 1 is as defined in formula nimVQSOffi ^ SMISATOViK ^ FLIMMWKVMS ^ JTXB 344 m3 ^ 6SS2 ^ imMV3QVE018%! ^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^ " 1, 2, 3, 4, 5, 6, 8, 9, 10, 11, 11, 11, 11, g &lt; &gt; ypoty3- · .--------------------------- '.............. ¾¾¾ 6 6msmsOTOTOTOTOTOTOTOTOTOTOTOTOTOTOTOTOTOTOTOTOTOTOTOTOTOTOTOTOTOTOTOTOTOTOTOTOTOTOTOTOTOTOTOTOTOTOT Ke Ke Ke Ke Ke Ke Ke Ke Ke Ke Ke Ke Ke Ke Ke Ke Ke Ke Ke Ke Ke Ke Ke Ke Ke Ke Ke Ke Ke - * * * --------------------------- 300 SW £ «WS9 - * EOOeB5lIlOS <JÊ3 ----- 308 ) 90883800830 ----------------------------------------------------------------------------------------------------------------------------------------------------------------- The invention relates to a process for the preparation of a medicament for the preparation of a medicament. ......... 391 5/23 FIGURE 3 Example 2 Exon 1 Stamping trimer Formation of the triple alpha helix BiPji1 3 'Q K? , K St 1 V Μ A S K Ble V37V Η Π F Π Π 51 51 51 51 51 Ϊ Ϊ! 0 8 H 3 U 'C l l 6/23 FIGURE 4 Fosition Tetraneotin brnaana V Tetranectin of aminol; Protein of cart. of bovine R Protein of carton, of bobarão § Consensus at 3 f g a h c V M T K tt F S B l K s a ii D ΐ V S s K M F & B b K ® E W D V ít V K E K D G O L K T E V K K S s K 3 R D E? X »R 13 K I D K I * deviant. b day c b k iHOÍVilliKSKÍillSítVCllÍ l w κ x v s ϋ s. κ and ϋ a &amp; lq τ v e l ÍS ÍS V V V V 88 S S S S S S 15 15 15 15 15! EXAMPLES 7/23 FIGURE 5 FIGURE 5 FIGURE 5 FIGURE 5 FIGURE 5 FIGURE 5 FIGURE 5 FIGURE 5 FIGURE 5 FIGURE 5 FIGURE 5 FIGURE 5 FIGURE 5 FIGURE 5 FIGURE 5 FIGURE 5 FIGURE 5 FIGURE 5 FIGURE 5 FIGURE 5 FIGURE 5 FIG. h 5 δ K? ϊ tf L Mati ^ o ^ ssTAtAeMtoGmTC ^ AracesK ^ cATCfiam ^ csOTK ^^ raAABAcmxTAcrassMáSec & rescceTTO twe f 6 VBP 8 BT ϊ K Eí VK, fc κ X β BKB «I p 5» <SORL Ϊ PA SOTTt! 3SISt'C, raGTOAtACCATT! 3ASiM3XCríM ^ caASÃTTCMas ^ dfôÈ0aarft: rcCC ^ OCTCaCSaaaaaaaaaaaaaaaaaaaaaaaaa * I. Tf fc VLRL «S β S Gv» Asa & ACT®MssTmaBTAm'TOTcmcTa <TATimAssasGirrftci, and, TTSATcmTeT'Ti''tiAG'w, eXisGeSSeSSssssssssssssssssssssssssssssssssssssssssssssssssssssssssssssssssssssssssssssssssssssssssssssssssssssssssssssssssssssssssssssssssssssssssssssssssssssssssssssssssssssssssssssssssssssssssssssssssssssssssssssssssssssssssssssssssssssssssssssssssssssssssssssssssssssssssssssssssssssssssssssssssssssssssssssssssssssssssssssssssssssssssssssssssssssssssssssssssssssssssssssssssssssssssssssssssssssssssssssssssssssssssssssssssssssssssssssssssssssssssssssssssssssssssssssssssssssssssssssssssssssss jjcaajocy y v s p q s s i is 3 and JR x 't, Jt x 1' s 'v' and v r s r x f $ & Agata et al., and the present invention relates to a method for the preparation of a compound of formula (I): wherein R 1, R 2, R 3 and R 4 are as defined above. ; tt; sgef &lt; tb &gt; &lt; tb &gt; S P V V 0 s F ir o H ti S K 8 Y B 9 L x e B ΐ ΐ ΐ ΐ ΐ ΐ ΐ ΐ ΐ ΐ ΐ ΐ K K K K K K K K K K K K K K K K K K K K K K K K K K K K K K K K K K K K g g g g g g g g g g g g g g g g g g g g g. why? 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 9, 10, 9, 10, 10, 10, 10, In addition, (i.e. s? &amp; The results are shown in Table 1 and Table 2 shows the results of the analysis of the results obtained in the literature. &amp; 'H' '' '' '' '' '' '' '' '' J A A B A C B S C C S C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C C X 1 'Λ S D L R Q 9 L l. The title compound was prepared from the title compound as a white solid, mp 218-221øC. 1 H NMR (CDCl 3):? βXXXCC VL · 8 3 FXV and P &amp; 8 '' '' '' '' '' '' '' '' '' '' '' '' '' '' '' '' '' '' '' '' '' '' '' '' '' '' '' '' '' ' FIG. 6 shows a cross-sectional view of a cross-sectional view of a cross-sectional view of a cross-sectional view of a cross-sectional view of a cross-sectional view of a cross-sectional view. cRacatmTCccTCtMjÍaÍÍA ^ tfrism.MCTO T? pjX »Ot®I o a h a η h a s s q j f v k t & t s k t r hr ^ WKJ ^ TATMftyftTCK ^ TCGcftTACCaTIcTTy ^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^ . S Ο T I B K V K A Κ I <J D K £ β I 1 »P 3 Q Q E L · í. Λ t-AcKixOAíirccAaTUACíCCÈTTOMaRTCTCAAAacrÃAíiAT TCAMasraÃGôíLtsQGTftTticeAcc-EGficCaijcsífiâtcTcS & Tíi.t if CT-3 ríGccâ read ST 33 0 D 1 S 0 0 KS II I Ϊ $ Γ h LK VL 1. R F S β β i3CXM.CASCWaA8RTOG®CtiT6CTTÍ'OTCTBACrACMm 'rCMMaOM'ICTACT ™' CMrrmTGraAESftCj: imTOTITOGM'CÇft Ssitt av 'ÍKCMOCMSáíJiyiGaArGTgítsaace-aiSiSgiggsgjAeetggísacegirísiggsiggsgigiggiggigiggiggagagaggsg Ϊ ES a 0 «e 2 3 V Ϊ Q SW SW X &gt; R v j D D t, A ΐ V V 'V D V 1 s r &gt; and β s "CAtíie ^ ciScaRgtttsssggeAesgffeítgggaAaaosgíUaaacctaaagasiaoELgaoaaefigggAoagcgtqa ^ KtcxiACCttcatuíasyutg yYSQRãOSAfiSKCliNtiKtilitltlÍillilV ^ TârESXl, egcgas £ ragnt; cggoccSgtgacficagsagt: EctgggataaccEggaaAagç (agacagaetgç catgaggca5ígagatí (agcaa &lt; qatctggagga3» £ δ t β PVT C and ϊ VB yl LSKSTE 3 1 * 0 K!?.! - S It &quot; Et t, s' is gísaagsceasggtgesgcccfeaeetggãcgaítkceasaagaagfeggeaggaggagatjígasstfitaeageeagâagesggasiícgctgsgsaea V JC AKV 0 Β γ L n B g ο K κ 'GSE' KLV Η 0 KVSP 1 KA sagotacaKgaggBaáogcgíiaagâasotgaacgag ^ tgEaasag & agatgsgeeeaatgggígaggagstacgagftaegígirgeaígcíoatgtg S I C R and Λ 6 ts 3 EE.? 2 K Si Si Si Si Si Si Si Si Si Si Si Λ Λ Λ Λ Λ Λ Λ Λ Λ Λ Λ Λ Λ 3 3 3 3 3 3 3 3 3 3 3 3 3 3 3 3 3 3 3 3 3 3 3 3 3 3 3 3 3 3 3 3 3 3 3 3 3 3 3 3 3 3 3 3 3 3 3 3 3 3 3 3 3 3 3 3 3 3. Ccg sgaa and ctgsg cg cgc teagogagaa ggeeaagoccgcgetegaçgacotscseeaaggcc tj «t gooe L A Κ Y H A Jf A T S». L · S T 1 ff K K A! to L h V gtgatggagaLg ct ttaagg t cag-c k 11 c ky ayagctct c gaggag t acfcc t sagasga tc that the ecagTM ^ MÍSCftPxIC ^ TgAATrCC ^ ATCC νΐ> «νκν Ρ] 8λΙ> 8 8ν« ΚΚύΚΤβ S! í ? (7'^ ^τττ BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS BS FIG. 7 is a cross-sectional view of a cross-sectional view of a cross-sectional view of a cross-sectional view of a cross-sectional view of a cross-sectional view of a cross- ^ ^ ^ ^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^ " t> VV ST? J <P 3 S x. £ ST ST -r L Λ QSV EI L Sf 8 QOA &quot; LQTV $ L 0 * TSV »3TCUV ^ C3 ^ GATG: meAaGftC <WT: AAC ^ ÇK; CaTClX3e * eACCC ^ CCCftG ^ B 'i' HX - JAOSGftTCCga CffaACíTcn-cxTcagâígrcccctífgsatcgagrtgaa ^ CCT ^ CôíiGÇíFt ^ Êeíítsawtfftírcçcâ ^ ageeáíjgfe ^ íiejAgrjc KOA ^ íJfíPPgÉÍPffSfíV ^ DLAÍVyvOVÍí / CDSGR * tatyVi ^ v &lt; & fft: ttg, A8F ^ ctt3 &lt; fôGBS9QP8 &amp; Ga & tttitity (&quot; Ggttgw &quot;) &quot; &quot; FF £ GS & L £ X is LXLXLL · υ KW ΰ 2 VT £ TFSX 'L and ^ SAS &gt; CAI (CbGffgccctgtg * oGtííggngçti; t ^ rt' 'oatgyMãã ^^ g æg ^ atfffgcct'g ^ GCI ^. (s): ## STR2 ## In the following examples, the term &quot; W &quot; and &quot; W &quot; and &quot; W &quot; (5), (5), (5), (5), (5) and (5) and (5). R K K Q jf V M S t V F: $ J √ V .8 λ »ϊ. A method according to any of the preceding claims, characterized in that the method of claim 1, wherein the compound of the formula (I): wherein X is an integer from 1 to 5, (c) &lt; tb &gt; &lt; tb &gt; JS &gt; QS h HRQX £? / 8? QSX? P?, GX 8% HDRA ???????????????????????????????????????????????????? The invention relates to a method for the preparation of a compound of the formula: ## STR1 ## in which R 1 is as defined in formula (I). p ¥ 2 U B l R? The results are shown in Table 1. The results are shown in Table 1. Table 1 - * ε K j, sr &amp; The compounds of the present invention may be prepared by the following methods: (a) reacting a compound of formula (I) with a compound of formula (I): ## STR1 ## in which: 2GCCATCXYOQCTSWAA ΥΙΐ'Β3νκν9 * 13ΆΙ Ν ΤΧΚΧ Ν Ν Ν Ν Ν Ν Ν Ν Ν Ν Ν Ν Ν Ν Ν Ν Ν Ν Ν Ν Ν Ν Ν Ν Ν Ν Ν Ν Ν Ν Ν Ν Ν Ν Ν Ν Ν Ν Ν Ν Ν Ν Ν Ν Ν Ν Ν Ν Ν Ν Ν Ν Ν Ν Ν Ν Ν Ν ATA-SCA-SCCJSAT- &lt; &numsp; &numsp; &numsp; &numsp; FIG. 8 (W7JBS Trip-JS-Apo-1-α) 143. Sa®5. ?. PB 'sa - (íruiil - ^ TCXCWcee ^ a ^ A3 ^ gsçoAXi ^ ErAt & GG ^ aCMRC.w &lt; SGTM crTCT &gt; ^ AAATAATiTKm'rAAqm'A.iu &amp;! M -' ArÍKT T7 projaot ^ r MG 3 Η Η Η Η Η Η Η Η Η Η "" "" "" "" "" "" "" "" "" "" "" "" "" "" "" "" "" "" "" "" "" "" "" Ϊ Ϊ Ϊ Ϊ Ϊ Ϊ Ϊ Ϊ Ϊ Ϊ Ϊ Ϊ Ϊ Ϊ Ϊ Ϊ Ϊ Ϊ Ϊ Ϊ Ϊ Ϊ Ϊ Ϊ Ϊ Ϊ Ϊ Ϊ Ϊ Ϊ Ϊ Ϊ Ϊ Ϊ Ϊ Ϊ Ϊ Ϊ Ϊ Ϊ Ϊ Ϊ Ϊ Ϊ Ϊ Ϊ Ϊ Ϊ Ϊ Ϊ Ϊ Ϊ Ϊ Ϊ , £ ®SOWaTate at &lt; M8Wv3A'iSíTTew5MÇTO (CiloscíWACTOOCTO iM ^ (x: Cak; ^ fttMTCGccoTOCTr aOGA3CKeC Vi3í5CC; rf &lt; 3CMiaceGTCTCCCT3 Ban al AAaasftTççcta.aagctccttsacaaetgggacagcgt gacetccaccttcegcâ.ajctg K-0 ϊ LK, Λ, U 'SSK WOSVTS Ij? aanaaetaggeeevgtgaccc acted ^ &amp; & amp gçKgcfcctgggataacctgga; aagBagasagagggCctg &amp; ggcagçagâtgagcaaggasetçsssíjgBg SSQLGPVT q "WDN i ϊ b &gt; B. and read TSG h SO ~ b 8 F t X) 1j and 33 gtgaaggccsaggtgcagrgfftaeceggscgaottoaagaaEraagtggcaggasgagatggagttctscccccsgaagsfcggagcagctgcgcgca VKAÍtv &quot; QPY 1 PO PO Ϊ Ϊ Ϊ Ϊ Ϊ Ϊ Ϊ Ϊ Ϊ Ϊ Ϊ Ϊ Ϊ Ϊ Ϊ Ϊ Ϊ Ϊ Ϊ Ϊ Ϊ Ϊ Ϊ Ϊ Ϊ Ϊ Ϊ Ϊ Ϊ Ϊ Ϊ Ϊ Ϊ Ϊ Ϊ Ϊ Ϊ Ϊ Ϊ Ϊ Ϊ Ϊ Ϊ Ϊ Μ 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5. To I. · 0 S K L 3 P i. 3 33 33 M R 0 R A a A Η V 'Becacgetaís ^^ catcfcggeeaiwLaaagegacgagctsc ^ ccag ^ ottggeegcgcgcctcgaggctcteaaggKjijaGggcggcíjccaçia DAÍ. R T H 8 A P Y 8 D S L · AA · RI · A · K · V K 6 8 A K ctggccgagtancacggcaAggscsccgagcaccPgag-acsctcsgcgagaaggneaagnncgcgcfccgaggaoctoegoaaaggcctgctgeee t · A B ϊ Η A T T B H i> 3 T b S tt K A K 3? A 'L E D Ϊ, R Q β tj 8? (A) (b) (b) (b) (b) (b) (b) (b) (c) t < S A J E E ¥ T K κ 'x, i * t ς evos κΐϊϊη; SACO (SOCIETY) (SOCIETY), SOCIETY (SOCIETY), SOCIETY (SOCIETY), SOCIETY, SOCIETY, SOCIETY, SOCIETY, SOCIETY, SOCIETY, ! TmremGgMO TW '' 3 'mcca' i- tacenv) -p83 '* 11/23 to FIGURE 9 ρΐ7 HSFSs Cys-Ape I -. &lt; Pvair) -aATC R7CCCgÍa ^ »awAft7AC TACftCTm ^ ^ ^ aRaacac CTm'mcfacTa ( .; Ta ^ ^ ^ f is'ETO laughing-i-tTJÍfttsAit &lt; yaÍSÍil'rft, i:? * T .mu.l: otr.òe CATATG * r ksshh.sk nBeexse TCGCATCACttXCftCCAICAQG ^ ^ ^ KTCM CGAGiWTAOa steak I Baitsi District eOTCSATCTiiatgsSoeeecoaaijagcpcctgygaucgastgaeggacetggcBaiísgçgtaogtçigatgcgct- .uaaMysnagoaasasagac CV CG &amp;? f S WD Ά The VK Ώ Ij ϊ ϊ v v 't &quot;! v LKDSG RB-tacangf.essaqttCgaHgqat .cagcattggjfaaaacagci.aaaSciâaíiscSCCtÇgíloaasCgggaoagcgtgacctccaccttçagijsÃgctg KVSQP' G * eAXcCIICQI.HiKtiI.O'B 'SVTSTP8Kí , Which is shown in Figs. 1a and 2b. Fig. 1 is a cross-sectional view of the Fig. .GtgaaggcGasggíígg cetA.acctggaegaettcicagaagaagfcggoaggaggagatggsgctcí-KS ^ .H ^ ^ gccagaaggtgfiegccgc segiÍSre '* V' AKVQP tJ 'BF &lt; 3 * K' OBSHei.XROXV *? '* G 3iKJk .gctCGaéK3agg3Sg' gcgcoasa.KgctGoaegagctgcaag 'gsagoí: * gíigoBKact3g5cía g3a3acgegc. A method according to any of claims 1 to 4, characterized in that: GSBBR 'RA δ Λ BV trans-BA ígfgotgagcRBgcstítggpcccctacagBgasgagstgogocagcgoBSggocgB.gígacttgatjaçtçtosâggagfiacggcggngccasa LRTBí- tfEDEI E1 / TCR RLAARLEABKSNGGAE ctggcGgagtsccacgcGaagGanHGsgagcsectGagcaceçieegfBgisgaiisgeeaagBecgcgQtcgaggaccuccgccaaggectgctgoc * I, A! * 3yHftK AT'BHÍ1STJjSSICAKPAIi2 »l! Ílíjei, gtgctggagagctteaaggtcagctteotgsgegeti ^ lP ^^ cgaggsgcacactaagasgcttíftBcaeeeagTAí rGCBÍgCíTaAjOTGCGKrcc V L κ 7 iS 6 'V g i', SA Ij £ S ϊ T κ B 1 Η X a STOP sphr Hiadlií KcoáIÊGCTGCTaacAAAacccGAssçsaMoar ^ wm ^ aitíCCTGCCAccGCTeAtStayTecTtaTACCTCTS-íG CCACraCTGÍSSCCTCXJ ^ CGGGTCTTGíAGGGOKXTTISCWAMGGAGGSACTATATCCGBT- ECORV! -pBE326. FIG. 10a FIG. 10a. FIG. s * Rm - '<w> ii> The present invention relates to a method for the preparation of a medicament for the treatment of a condition in which the medicament is administered to a patient in the form of a medicament. pínjwPtfci 'M <3 S K B H H K »a S I Q S 5. 8 P a T Σ 9 í> TA k i V KK 3 8 The ATACÃTAi Α κ ^ oc ^ TCAC &gt;; ^ KacraTCAm30 saATc: ACC aAiçcA®B'iíWOTCT ^ '^ AaccAC .cíe A3CC c ^% ^ ^ TW5m5ATaccAW3fir GCI: s Bgl II:! - 4 :: 1 Τ 'DVV Η ΐ KKPES 3 * KSA J D D Q Q Q VA VA VA VA VA VA VA VA VA VA 0 0 0 0 0 0 0 0 0 0 0 0 0 0 a a a a a a a a a a a a a a a a a a a a a a a a a a a a a a a a a a a a a a a a amp amp amp amp amp amp amp amp amp amp amp. (1) and (2), and (2) and (3) and (4) and (4) and (4). ? - £ p p p p ΰ ΰ ΰ ΰ ΰ ϊ ϊ ϊ ϊ ϊ ϊ ϊ ϊ ϊ ϊ ϊ ϊ ϊ ϊ ϊ ϊ ϊ ϊ ϊ ϊ ϊ ϊ ϊ ϊ ϊ ϊ ϊ ϊ ϊ ϊ ϊ ϊ ϊ ϊ ϊ ϊ ϊ ϊ ϊ ϊ ϊ ϊ ϊ ϊ ϊ ϊ ϊ ϊ ϊ ϊ ϊ ϊ ϊ ϊ ϊ ϊ ϊ ϊ ϊ ϊ ϊ ϊ ϊ ϊ ϊ ϊ ϊ ϊ ϊ ϊ ϊ ϊ ϊ (1) (1) (2) (1) (1) (1) (1) (2) 'D II £ SX Β VS &lt; 3 G · MQXHSI t read it eszqaxqgcoaiiqiztgtzãgccctsGSítgg & àCíaçttzzí &amp; sszGAxgi-.ggcsggjiçigagatggãgcccMccgccsgaasçrtgyftgccgci.gcgcíicií v y;.. jc &quot; v &lt; j i i odf the &quot; K jc wqaa KjítysoKVífyjiK GA C ^ c ^ c ^ c ^ cgcgiTgce & ga: cgagc fcgcaàga faag * "yAOocCA c ^ Ε / s λ qs õ / s / / / / / / í í í í í í í í í í í í í í í í í í í í í í í í í í í í í í í í í í í í í í í í í t t t t t t t t t t t t t t t t t t t t t t t aa ig aa aa aa aa ^ ^ ^ ^ ^ ^ ^ ^ ^ ^ ^ ^ ^ ^ ^ ^ ^ ^ ϋ ϋ ϋ ϋ ϋ ϋ ϋ ϋ ϋ ϋ ϋ ϋ ϋ ϋ ϋ ϋ ϋ ϋ ϋ ϋ ϋ ϋ ϋ ϋ ϋ ϋ ϋ ϋ ϋ ϋ ϋ ϋ ϋ ϋ ϋ ϋ ϋ ϋ ϋ ϋ ϋ::::::: ΙΑίι ' ϊ Α ΐ ΐ Λ Λ Λ Λ Λ Λ Λ Λ Λ Λ Λ Λ Λ Λ Λ Λ Λ Λ Λ Λ Λ Λ Λ Λ Λ Λ Λ Λ Λ Λ Λ Λ Λ Λ Λ Λ Λ Λ Λ Λ Λ Λ Λ I B F F. &Quot; V B? I &gt; SA, ES γ TKK í Γ Γ Γ Γ Γ Γ Γ Γ Γ Γ Γ Γ Γ Γ Γ Γ Γ Γ Γ Γ Γ Γ Γ Γ Γ Γ Γ Γ Γ Γ Γ Γ Γ Γ Γ Γ Γ Γ Γ Γ Γ Γ Γ Γ Γ Γ Γ Γ Γ Γ Γ Γ Γ Γ Γ Γ Γ Γ Γ Γ Γ Γ SI SI SI SI SI SI SI SI SI SI SI SI SI SI : - 83-35 3. Figure 10B 13/23 I - AiflíA JsWmS-ffmiíIi -SMCXCmTCIX IBAA ^ ^ ^ raAhTACEíÍWJftCTAÍAaSaAOACCAJKAPBfflrmÍglCTJijAAATAA TrtmTCAACÍ trAAaiÍACeAviaT-T7-t ?; protjfot NRKHaKHSSGSSSOSSSOF eTEePjgxjK K-SS IVSA AW / Amitil raKSTCACtoroCtoKAa ^ ^ rM &gt; TCroA0TOtovt. Β ΐ ΐ ΐ ΐ ΐ I. I. I. I. I. I. I. I. I. I. I. I. I. I. I. I. I. I. I. I. I. I. I. Β Β Β Β Β Β Β Β Β Β Β Β Β Β Β Β Β Β Β Β Β Β Β Β Β Β Β Β Β Β Β Β Β Β Β Β Β Β Β Β Β Β Β Β Β Β. A &lt; 3a &s; &lt; 3a &gt; &lt; / RTI &gt; &lt; / RTI &gt;; &gt; &gt; &gt; &gt; &gt; &gt; &gt; &gt; &gt; &gt; L A T V f V V V V V V V V V V V V V V V V V V V V V V V V V V V V V V V V V V V V V V V V V V V V V V V V V V V V V V V V V V V V V V V V V V V V V V V V V V V V V V V V V V V V V V V V V V V V V V V V V V V V V V V V V V V V V V V V V V V V V V V V V V V V V V V V V V V V V V V V V V V V V V V V V V V V V V V V V V V V V V V V V V V V V V V V V V V V- I &gt; 'HCSVTS ΐ £ K P · l cgftgaacsgctcggecctgtgacicesggBguctgggatsacctggftaâaggagaeagagíggretgaggíaggagAigagcaAggatetggasçísg jt SQ 3- (PV 5 and y 1 h SDK K 5C STS δ Ιι &amp;? Ό SBMS S. L V EE gtga.aggílsaiíggtgiMgocctAcctggaegaef.tisragaagaagtggoarjgaggagatggagctcfcaccgaesaaaagcâgagccgev.gogagoa KSXTiOSVt.DDFQKKifQ KXMSÍiíKQKVBíi. RA-gasítftwaagegggíig ^ .- iji.OagaRgíiííidígagcítgeaagAgAAgfetgagc csrtgsgojagsaáfaiSíígíffÃaffgiigggsijBgcocRtgtg β'ΐ.ςΐ''ϊΕβ ** &lt; ϊΚϊ · ΗΒΙ.ΟΒΚ: ϊ.ΒίΙι &lt; 3ΒΒ 'ΙΙ'ΒϊΙΒ.ΒΛ' ΐν gaegcggtsggcacgíatctggcccoctscagogaegagetgcgncagcgettgaccgegcçecttgagactetcaaggagaacggesgogeoaga P AI aTa & gt .l.í , P P Β Β Β;;;;; SB SB SB SB SB SB SB SB ΐ ΐ ΐ ΐ ΐ ΐ ΐ ΐ ΐ ΐ ΐ ΐ ΐ ΐ ΐ ΐ ΐ ΐ ΐ ΐ ΐ ΐ ΐ ΐ ΐ ΐ ΐ ΐ ΐ ΐ ΐ ΐ ΐ ΐ ΐ ΐ ΐ ΐ ΐ ΐ ΐ ΐ ΐ ΐ ΐ ΐ ΐ ΐ ΐ ΐ ΐ ΐ ΐ ΐ ΐ ΐ ΐ ΐ ΐ ΐ ΐ - P gegoTggagasctfceaaggCcagctÉccfcgagcgCtCtcgaggagtaoa ^ raagaãgctcaacav VCaaTMTA ^ aMISAKWK ^ luATOCfiGCTCrXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXX ; v ^ I: ¢ τ ΓBΓΐ'KCiccACCS &lt; IXIíBKCcra ¾AACi.Mfiϊ'crros ^; ^ 3 ¾w ΤΤΤ1ΤΤ®: ϊ 'ΑΑ ι0Ι3ΑΗ (5ίΜΤΑΪ ϊ1χ · Ο2 & Τ IficoRVi-f-AI2 *!'. Figure 10c): Arip-A-rn-λρο Al-final - Amp, PBS328 - <mix t BATCT03Ascccac ^ ARTTAft ^ AOi ^ cA (^ M'à ^ c> gmicMiRsa89rmsx ΗΗ ΗΗ ΗΗ ΗΗ ΗΗ ΗΗ ΗΗ ΗΗ ΗΗ ΗΗ ΗΗ ΗΗ ΗΗ ΗΗ ΗΗ ΗΗ ΗΗ ΗΗ ΗΗ ΗΗ ΗΗ ΗΗ ΗΗ ΗΗ ΗΗ ϊ ϊ ϊ ϊ ϊ ϊ ϊ ϊ ϊ ϊ ϊ ϊ ϊ ϊ ϊ ϊ ϊ ϊ ϊ ϊ ϊ ϊ ϊ ϊ ϊ ϊ ϊ ϊ ϊ ϊ ϊ ϊ ϊ ϊ ϊ . ο τ a to g va * τ r .. xxsgealg v $ g ^ ri ^^ aKjAAacaramTOwacAA saAQCTcMiaacoaMMaAoswce açsci ^ ^ ^ Awieaccctáerai wtKAeíyi Mee ^ H ^ ccwcfifíACíJfíícicocw eara iAÇSieaijstjaíjg-svSMceooffooagagceenr.íjggac.ogagtgaagçiacotggooaaçsKgCRccsgsstgtgotoaaaganaacgscAgaga.a Κ &quot; X ο »g 3 »* 80 80 80 80 80 80 80 80 80 80 3 3 3 3 3 3 3 3 3 3 3 3 3 3 3 3 ϊ 3 3 3 ϊ ϊ ϊ ϊ ϊ ϊ ϊ ϊ ϊ ϊ ϊ ϊ ϊ ϊ ϊ ϊ ϊ ϊ ϊ ϊ ϊ ϊ ϊ ϊ ϊ ϊ ϊ ϊ ϊ ϊ ϊ ϊ ϊ i3 X 0 1 &gt; HEKX, i &gt; ÈJ MDSV Ϊ S Ϊ F 3 K I. agtigaaaagetcggíectgtgacccaggagttcfcgggâtaaiweçgaaã & ggagaeagsgggeesçaggcaggagabjagoaaggstctgg & qqag eB .. Q'l, GPVíO.Bf 'D' i KKSTBeúàQSMSKDtBe gÇgaaggceaâggSgeagooeeaoçEggacgaeEtcc &amp; gsagaagíggeaggaggagatggagçíctaccgccagaaíggiiggagccgc ucgegc ^ * VK V AX QÍYl.íIOfaKKWÕSSS S iTSOX VEPi.RA 9agcUccaac (agc (g2g2g'2gccagKaçctct: acgagt; t.gc9agâjgaag2tg3gcccacr.gggag »^ ggagAtgogsgascgc3C3c egescat Tg (5 I &gt; 0 's'' A R Ç X I> Η Η ΰ ΰ ΰ ΰ ΰ ΰ ΰ 5 Η Η Η Η Η Η Η Η Η. R 3 R 7 &gt; R A Η Vg c cgcgcCgcgGa2gcaC2 g í í í::::::::::::::: oa oa oa oa D D D D D D D D D D D D D D D D D D D D D D D D D D D D D D D D. 3 L A? Ϊ Ϊ L L L L L L L L L L L L L L L L L L L L L L L L L L L L L L L L L L L L L L L L L L L L L L L L L L L L L L L L L L L L L L L L L L L L L L L L L L L L L L L L? A L 'B D L S Q S L L R ghgctggagagctKsaaggtcagcttcctgagcgotStcgagçsgtacacfAagaagiiCSÃaasoooagiCAATAAKvrts ^^ JgO.ewíCOeííSTUcrAli V T < E r »R κ V E? LSALEE ϊ ΐ Κ Ϊ S x L to stoi SiaiMIÍ SioF.l CAARSOTiKwsAíWsAaxs siitíSRi ^ ^ ^ íXBtriacwcia gSTitóeeiwa fnTWAWTASwwcTwwc ^ ^ ^ c ^ s ^^ eCTejwMKx aiSeiWíGAsscis TCTTTTiieTGAAAeeAaSWSCrATSSOCSSR- (E20RV! -1 * 832 '. FIGURE lOd 15/23 I- T7HS-kA-K1A-K1SA-W * &lt; pBP.323- fgvitll -GA ^ TQGA-J ^ CC ^ AAAA ^ CFAA ^ AC ^ ATAC ^ RTI ID = 0.0 &gt; ACT &lt; / RTI &gt; &lt; ATCA &lt; RTI ID = 0.0 &gt; ACK &lt; YV33? K * FKfiLK? RX, BT & JLQBVJIJK? QSQOAI QTVSI? AA? T5TTG? GJUCAC? AAGASSTriSftG3AGvrrCA? W ^ C ^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^ " czGg ACfg ^ ^ ^ ^ w IIIR àãáLctáéLgiQaLgg al ^ &amp; ^ tat cgefiacctri Lamti ^ c ^ ^ ^ tca aiaAcaaaqaaattaftaG K ^ t ^ will cfqDEÍPQSpíjORVKDLftTVyVJDV LKinSGRO tatgvÇjtçcGagc.ttgaaggcteGiiocttgygíiítaac cíaaacctaságctccttga.CÃactgggacagoçtySCCtOC cctt ^ ε-cAgcaâgctg qf v? egsa ii α κ i q ij nl l jc r &gt;.? Ic PRRS fwusv τ i .. '^ ^ gaacAgcir.íggccctgtgaíx aggagttctgggataacctggaaaaggagacags ^ gg ^ ^ Ra ctgaggcaggagatgagcaaggatctggraggag tCPVTQKFH KÍtiKK ^ ^ ^ KâtiROSHSlCOlie.S gtgaacgccftaggtgcagccctaGctggaçgacttccagaaga gtssjcacigaggag & tggagehctâCcgCwftgaa ^ ^ etggaíícccíCtgcgrggca VSCAKVO ti) t &gt; .D ^ OR'RÍtâB8MlB !! (Í ¥ Í »QKVB ^ IIRA gagctccAagagggcrcfGgcgcc &amp; gaagccgcacíjâgEtgcaagagaagctgagcccactggsfcgagg &amp; gatgcgcgaccgcgcgcgcçfCCcatgtg ETICS &lt; 3AEÇKfcH8: & QB £ i SPLGBB H £ BRARAHV gAcgcgctgcgcacgcahetggGCCddtacagígôegâgíifcgcgccagcgcEfegeccgcgGgFeefctgaggGtctcaagçagaacsfgcgííCÍgeeaga ΡΑΐιΚτκϊ, Αργε'ΐ &gt; ϊ: 1ι .alpha.,. &lt; / RTI &gt; jf &quot; s. S g ^ r ^ SSCsSf fcasoacgecaaggdçaççgaeeAtCt qeâegGfce ^ & & amp gcgaqsagqcreaâgccegcaefceqaqqauetceífce; agtfeefcgetgcGís IjÃÍÍYKARATtíHliSTIití'EKAKÍ ALBI *} l &gt; RGÍíIfÍ: P ^ ^ gítSgag sgccr.caaggtçaguçcçcitgAgcgufcçtçgsggagtycaçtaagaagGtcaacaçGcagTaATAMcyraMritpGíkTC GGCrOCTAA V ^ X, δ S? K V S P l 's a L £ E Y T K K T &lt; Ϊ Q ST & P ΚϊλΛϊΙΪ fte * aa CAM5tCOTAAaGΠAAGΓTGAGT ^ Π (^ TGCCTGrCACCGCTC¾'βCTΰAGCJ ^ AT ^ OVCTA ^ AΐA ^ CCrtrΓCIGíXA'r: CGCΓιm ^ ¾C ^ CTf ^ AACaGOTCTm¾ϊ3 <3β TnTl * KCISÁAAGGAGd & eTAyATCí & AT- (ftcofcv FIG. 10 is a cross-sectional view of a cross-sectional view of a cross-sectional view of a cross-sectional view of a cross-sectional view of a catheter. &lt; tb &gt; &lt; tb &gt; &lt; tb &gt; &lt; tb &gt; &lt; tb &gt; M &gt; and gBTRCQaAgCCftCi ^ 0CCA3 ^ OCGOM.i: 'ARGMTGT.WtTgcc agi li φ φ & & K K p p p p p p p p p p RE RE RE RE RE RE RE RE RE Λ Λ Λ Λ Λ Λ Λ Λ Λ Λ Λ Λ Λ Λ Λ Λ Λ Λ. TBXSB ÃaOACAWSÍOmaAaaASCTCM ^ &amp; ^ WearMaSftrAOfKTSOiXaWASSTOíÍeCWaCTSAM ÇASCMeCCCrMICAGACGaTCTffiCM 6 'k Κϊ AfciWtotioeKRÍiCTrataqiffnÍUtagMiBceccooPitaacceetgaaaeetMwtqaajigaccCcMccwotBtataagiagesatactcaMgaoegcgBc K G S VS μ &quot; jf DSPPQS' 'to VKD Ϊ1 kt V ^ ϊ the VDVLSDSG &amp; ^ c ^ ii ^ tcciCagtíít gt apggctí ^ ^ ^ gacttgfigâíLííscgacSaíiacctaBãgctccttgaoaactgggatíÈÍCJcgtgacctccaçít sa ^ ^ c ^ RD asqí ΐ VSQ 'ESSRLGXQ KL i K 1 · DKHDSVSSTPSKL egagaacagcScKigcííatgjtgaepfiasgagtfcstgsqacaaect.Hg.íaaíiggaçracagagggcctgaggcs.ggoSAtgagtíaaggatetggK.?. ggag R.B 0 5, S s V Ϊ QB! W 'c to L 5 K K 1 S <3 h! T Q B' 8 A D l. E B gtgaaggsaaaaÍiUicagcWítsiiftlÍggsogaottooagaasaaçjtgflcagçfsggagfitggageCosaoogccagasggtggagBcgataegggca νΚΑίί ν0ΡΥϋϊ &gt; ΒΪ · ΰΚί 'ΰΚ' ΜΒΙ.ΪΚ 65ίνΚΡΙ.ΚΑ qagetcííaaqangggg rgcigccagBagcitgaaegagetgoeagagaagctgagiiceagf.gggcgaggssatgcgcgaMKfCJijgisçregctrcatgtg B ^ G 0 · V S R &lt; / K E L I 8 0 8 K & S P -I. S E R E R E R E R E R E H V geegcgstgcgoacgcatotagiÍoiiijirtacagcgaegasctgMtí ^ cagígiíttgfniJsasngcotegõggoEcMeaggagsauggnggegecaga o'aí.'rt'híarvsdbi »sori. to Λ, R. 1 E A L X S N <3 S Λ R gg gg «« «« e e e e e e e e e e e e e e e e e e e e e e e e age age age age age age age age age age age ac ac ac ac ac Ϊ Ϊ Ϊ Ϊ. JS Y K A K A T B H i> STL a BK Ά KP ή h SO 1 RQ β I. LS »gtgaessa <p ^ ttcaaggi: s« 9CKfcectgeg ^ iotc6tyae9 »9 * (1) (1) (1) (2) (1) (1) (2) (1) (2) (3) bcv-pflfii 2 a - 17/23 FIGURE 10 pT70S Txip-Λ Γ :; Λρ * Ai-final - - tog * .PBHSis- ρρνρΐι} -GArc; AG: vi: ix: GCAAAAyiaATACOA? ftcaCr; AA; ^: ACKNOWLEDGMENTS ACKNOWLEDGMENTS ACKNOWLEDGMENTS ACKNOWLEDGMENTS ACKNOWLEDGMENTS ACKNOWLEDGMENTS ACKNOWLEDGMENTS ACKNOWLEDGMENTS ACKNOWLEDGMENTS ACKNOWLEDGMENTS ACKNOWLEDGMENTS ACKNOWLEDGMENTS ACKNOWLEDGMENTS ACKNOWLEDGMENTS ACKNOWLEDGMENTS ACKNOWLEDGMENTS ACKNOWLEDGMENTS ACKNOWLEDGMENTS ACKNOWLEDGMENTS ACKNOWLEDGMENTS ACKNOWLEDGMENTS ACKNOWLEDGMENT ACKNOWLEDGMENTS TOcgev TrogTAgivs ^ ^ ^ TCA CAV TAOftTeTCCTastÃccá ^ ^ ^ mcgftaccx mgccgm ^ ffiftTTOiVÍiÍAsgsg SsiTi Sj® Τ '¥ XS DVVtf RMFSELX5 tl, BTL!.' lQBVAi; {{IK8 0ai.GTVj;! i 'AAAI GMwrrorowutvfcvíí ^ ^ ^ iwmwcíi xveiAeAccemcrasABACxXGracRXíAieM ^ Saai SII AMtjaftACCVvRasy ^ AcaiOim »aacccgcgcagagcccefcgggaf.cgaqf cgaRqqacetqqccactgtstae 'tcjc; 3tgtqcl: gaaaqagBgcggc K' O 'T K v' Η Η K t> (1) (2) (1) (2) (1) (1) (2) (1) (1) (2). YV 3 0 * FK '<3 SAI »β KQ 1. DLK Tj Ií" 0 tf κ "θ SV TSTFfíKl · cqeg' ae 'BeLcggeectgtgacccaaaitgttat8S9WC« «ecca8« HMm8g «9i» «» g8gctaaggeacaagat983CM> g8afccfcg8a99a3 R KGfcSFVTCiEFHStS !, EK BT SS3 8.QKMSK3 «BE • gtgaaggw ^ aggtgcagoeetaeoigsasiSa iCfiaoagsa« aagt: ggcagg-iggagat; gge, gct; tasagacagaag3i; gge; scgca 'v ec &quot; ã κ v ο ρ γ ldpp ο κ κ «Aebhelira κ vei? l k λ gaget-BeaagagggagegogacagaagatgcacgageEgcasgagaagotgagccoactgggtgaggagatgcgcgaccgcgBgegcgcccacgtg S t. a S 0 A R Q S L L E 1 Q 3 Κ I, 8 P L S E E R S S κ S V gacgcgafcgcgcaí; gcatcCggcccc01; there: agagg; gagctgcgdcagcgcttggegigigggtgggotgtgaggagaadggcggcg; δ T H L A? Y S t> S I, R 0 S if Ã Λ E L S A L Κ B K3 S A K ecggccgagEaBoacgccaaggBsacsgagoaactgígacacagoaagogagsaggouaagotwgcgBeBgaggacctficgcsâggcistgatgBsc h A '0 and Η Λ Κ ϊ ϊ ϊ ϊ R R R R R R R R B B B B B B B B B B B. gtgBtggagagettcaaggtc-agBttBítgágBgBEuuçgagtiayEatiaciEaag.a-agetcaãdaBBeagTMTAA TTGAAtTCCflATCCfíÍTCTytrrAA ^ V 1 '(SFXV, SF 3 I &quot; 8 AL' 3 ϊ '1' κ K h HSQ iraw Ηίηώϊϊ'κΒίϊ (g ^ ^ ^ AAOÇCBMAASí KSAOTTSaa BTGmACC ^ ^ ^ TCA TSBaBWiTftJfCTftaOATAAftCXSíCTCCCRCrat FIGURE 10g pT-ne 'ϊΓ ΰ ΰ ΰ ΰ ΰ ΰ ΰ ΰ ΰ sinal ΰ ΰ ΰ ΰ ΰ ΰ ΰ ΰ ΰ ΰ ΰ ΰ ΰ ΰ ΰ ΰ ΰ ΰ ΰ sinal 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 : TJmgBG; iSA ^ cACWca «''TTTT A A A A A A A A A A A A A A A A A A A A A A A A A A A A A A H H H H H H H H H H H H H H H H H H H H H H H H H H H H H H H H H H H H H H H H H H H H H H H H H H H H H H H H H H H H H H H H H H H H H H H H H H H H H H H H H H H H H H H H H H H H H H H H H H H H H H H H H H H H H H H H H H H H H H H H H H H S? STS 2 P Ϊ 0 ϊ X Λ ΐ V AA AA ag ag ag ag ag ag ag ag ag ag ag ag ag ag ag ag ag ag ag ag ag ag ΐ ΐ ΐ ΐ ΐ ΐ ΐ ΐ ΐ ΐ ΐ ΐ ΐ ΐ ΐ ΐ ΐ ΐ ΐ ΐ ΐ ΐ ϊ ϊ ϊ ϊ ϊ ϊ ϊ ϊ ϊ ϊ ϊ ϊ ϊ ϊ ϊ ϊ ϊ ϊ , Ϊ́ϊ ΐϊ ΐϊ ΐϊ ϊ ϊ ϊ ϊ ϊ ϊ ϊ ϊ ϊ ϊ ϊ ϊ ϊ ϊ ϊ ϊ ϊ ϊ ϊ ϊ ϊ ϊ ϊ ϊ ϊ ϊ ϊ ϊ ϊ ϊ ϊ ϊ ϊ ϊ ϊ ϊ ϊ ϊ ϊ ϊ ϊ ϊ ϊ ϊ ϊ ϊ ϊ ϊ ϊ ϊ ϊ ϊ ϊ ϊ ϊ ϊ ϊ ϊ ϊ ϊ ϊ ϊ ϊ ϊ ϊ ϊ ϊ ϊ ϊ ϊ ϊ ϊ ϊ ϊ ϊ. ccM ^ v3 ^ rcc: ronWAi f (s) and (c) (c) and (b) (c) and (b), and (b) (c) and (b) 0 LATV ϊ VDV 1 SE &quot; * S &lt; 3 sgacfictatgtçiítocca.gtttqaaggetccgcctvSgsaAaacagotsUíicctaÃagctoiJttgasaaatgijgaoagcgtgaggtscagcttcagcEagaStí R t) XVS o FE and SALGKQL fl L · XLT; 0 Μ »t> S V Ί S Ϊ F S K L «gegascagctcggccotgtg.icccagçiagttctgçjgata.aectggKeaflggagacasagggcctgsggssggagatgasjaaaggatcbggaggsg R K ¢. Fig. SB gbg.xuggccaaggtgcagígrtacgtngaiígacttSíagaagaagtggcaggaggâgâtggagctctaccgCRagKaggtggagacgctssgaaaa V K ft X ·? The R-, D,,,, K K K K K K K K K K K K K K K K K K K K K K. gagt; t.cca & gigggcgcgcgccagr) / ig'gwgcacgag®lig &lt;? aagagaagiitgsgcorractgggcgn59ngKfct5c! 3c: gst: tggagftgg ^ cQcat; RAKAB &quot; v gacGcgcLgcs-dog ^ gBtctggcccootacagpaadgagatgGgfigagcgettggnngogggKGr.tgagggSfitÈaatigagafiogacggçgGasLgã H R ™ KI Ά p Ύ S the ELR g IR, a Λ k I &gt; BALKEHGGA it eeggccg * GT &lt; sQcaegceaRgeeeM: egagoat 'tg' 90 'eGate' gegi ' (3), (3), (4) and (4) and (4) and (4) and (4) and (4) and (4). ^ Ct <i3aa3sat »88ca« W8a ^ CcaacacccacffMTRRerfTtttMTOm «Aaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaa aaaaaaa FIGURE 10 FIGURE 10 FIGURE 10 FIGURE 10 FIGURE 10 FIGURE 10 FIGURE 10 FIGURE 10 FIGURE 10 FIGURE 10 FIGURE 10 FIGURE 10 FIGURE 10 FIGURE 10 FIGURE 10 FIGURE 10 FIGURE 10 FIGURE 10 FIGURE 10 FIGURE 10 FIGURE 10 FIGURE 10 FIGURE 10 FIGURE 10 FIGURE 10 FIGURE 10 FIGURE 10 FIGURE 10 FIGURE 10 FIGURE 10 A-A} x: - Amino Amino Acid Amino Acid Amino Acid Amino Acid Amino Acid Amino Acid Amino Acid Amino Acid Amino Acid Amino Acid Amino Acid Amino Acid Amino Acid Amino Acid Amino Acid Amino- pxcT ^ tcr M9BHea'JIIÍÍfesjiaaR <JVDSnSOVSDI AOS.OCPKP,? The compounds of the present invention may be prepared by the following methods: (a) Preparation of a compound of the formula: ## STR1 ## wherein R 1 is as defined in formula (I): ## STR4 ## ............. h 1 b or i> and v i? & (1) and (2) and (2), and (2), and (2), and (2) are the same as those in the case of the Tgga-sgga; (1) (1) (1) (1) (1) (1) (1) (2) (2) (2) (2) (a) (b) ;, * gO7tt5y0aaõsragcta3 oot: íâ33rct'i; cftsfacaâcS3g '^ íacagc cgacwt: wacci; tí.agi: ®a9íc (: 7 g ΐ SCF S £ Λ f 0 A &lt; 1 jii 5g Q L · k i LZ .V s' £ 'SV 3' 8 Ϊ 'Γ κ f / cgqgaacaiK wgssecesítgífflsíaírsagctctggjgõtaai-ijssfg-aaaaíEagasilgasrsíxíltg-ajjlícasfífapjratsfáswaafgatrcsrg-ag-gag · SECiiíPVrffSfWOfíiieEBrEeiAOSWSEOLES 9l &amp; Wneea * g8tgBiigtZ cUKXX &amp;! gacgactttía &amp;! TA8 * wg ^' WGG 'gatggiíffccct' cagee * * * * * * • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • tb> gt; gt; gt; gt; gt; gt; gt; gt; gt; gt; gt; gt; 'AAV jfacscgctgpxfcscgcafcctjiíccccctaGSSffgacífajccgcgccsjcgcttsfjrtxíSfiscsecSt ^ g-Si ^ rçtcasgjfstraacgjojfçrcyccaífe CA Λ Α "Γ II £ A i' f 5 &gt; 7 ff £ S c * R · i A 'SÍJItíKSSí SA 9 Ctjgccgagtafptracgccaaj ^ ccaíWSfagcaCírsatgcsctttCoj ^ cgasfaaggíCCSagcâcjcsctcgajgaCTCÍiccjccaajjíccEgccjifec IT AS k' MA 'ATEHA 8 T h SSSA * f PA t, S 0 · K K K £ Ρ Ρ Ρ Ρ Ρ Ρ Ρ Ρ Ρ Ρ Ρ Ρ Ρ Ρ Ρ Β Β Κ Κ Κ Κ Κ Κ Κ Κ Κ Κ Κ Κ Κ Κ Κ Κ Κ Κ Κ Κ Κ Κ Κ Κ Κ Κ Κ Κ «« e &gt; ccaa &quot; Aaicfasj &quot; cAgtxaatA &quot; AIPCWStMmBeaMetsaagaCTa &gt; v 1 · e g? κ √ √ √ √ √ √ √ √ √ √ √ √ √ √ √ √ √ √ √ √ √ √ √ √ √ √ √ √ √ √ √ √ √ √ √ √ √ √ √ √ √ √ √ √ 1 -Mu. FIGURE 11 I DíviPC + Trip-A-Ai ο, ο4 » DMPC + T rip-A-FN-A! j <or '0.0 · 0.- 0.4 · 0.2 · 0 · 0.4 mg / mi DMPC Seia Protein | Time (min) 21/23 FIGURE 12 Linkage of apo A-I and Trip-A-AI to immobilized oubillia Response Unidscies Τ Τ Τ Τ Τ Τ Τ Τ Τ FIG FIG FIG FIG FIG FIG FIG FIG FIG FIG FIG FIG FIG FIG FIG FIG FIG FIG FIG FIG FIG FIG FIG FIG FIG FIG 23/23 FIGURE 14 Comparison of the plasma concentration of Trip-A-I and Αρσ A-I for 2 days after 1 day ingestion, mean of 5 mice Relative plasma concentration. Time {hours}