TW202245833A - Pharmaceutical composition for subcutaneous injection comprising human hyaluronidase ph20 variant and drug - Google Patents

Abstract

A pharmaceutical composition comprising: (a)pembrolizumab; and (b)a PH20 variant comprising the amino acid sequence of any one of SEQ ID NOs:5 to 50.

Description

Pharmaceutical composition comprising human hyaluronidase PH20 variant and medicament for subcutaneous injection

The present invention relates to a pharmaceutical composition, comprising a human hyaluronidase PH20 variant with enhanced enzyme activity and thermostability and one or more drugs, and a method for treating diseases using the same.

The pharmaceutical composition according to the present invention can preferably be used for subcutaneous injection.

Drugs that should be given in high doses or multiple doses, especially drugs such as antibody drugs, are usually administered through intravenous injection, which takes about 90 minutes or more, and intravenous injection is accompanied by additional preparation procedures, so it is very difficult for patients and doctors. and medical staff is quite inconvenient, and additional costs will be incurred. In contrast, subcutaneous injection has the advantage of being able to administer immediately, but compared with intravenous injection, the absorption rate is relatively low, and when the injection volume is 3-5 ml or more, due to slow absorption, it is difficult to May cause swelling and pain. Therefore, subcutaneous injections of protein therapeutics are generally limited to small solution injections of 2 milliliters or less. However, when hyaluronidase is subcutaneously administered (or subcutaneously injected) together with therapeutic drugs, hyaluronic acid distributed in the extracellular matrix is hydrolyzed by the action of hyaluronidase, so the viscosity of the subcutaneous area decreases and the permeability of the substance increases, so , high doses or multiple doses of drugs can be easily delivered to the body.

There are six hyaluronidase genes in the human body: Hyal1, Hyal2, Hyal3, Hyal4, HyalPS1 and PH20/SPAM1. Hyal1 and Hyal2 are expressed in most tissues, and PH20/SPAM1 (hereinafter referred to as PH20) is expressed in sperm cell membrane and acrosomal membrane. HyalPS1 is not expressed because it is a pseudogene. PH20 is an enzyme (EC 3.2.1.35) that cuts the β-1,4 bond between N-acetylglucosamine and glucuronic acid, which are the sugars that make up hyaluronic acid. Human hyaluronidase PH20 has an optimal pH value of 5.5, but exhibits some activity even at pH 7-8, while other human hyaluronidases including Hyal1 have an optimal pH value of 3-4 and exhibit some activity at pH 7-8. rather weak activity. The pH value of the subcutaneous area in the human body is about 7.4, which is essentially neutral. Therefore, among various types of hyaluronidase, PH20 is widely used in clinical use. Examples of clinical use of PH20 include subcutaneous injection of antibody therapeutics, use as an eye relaxant and anesthesia additive in ophthalmic surgery, increase anticancer by hydrolyzing hyaluronic acid located in the extracellular matrix of tumor cells Therapeutic agents enter tumor cells and are used to facilitate resorption of body fluids and blood present in excess in tissues.

Meanwhile, currently commercially available PH20 is in the form of extracting from the testes of cattle or sheep. Examples include Amphadase® (bovine hyaluronidase) and Vitrase® (sheep hyaluronidase).

Bovine testicular hyaluronidase (BTH) was obtained by removing the message peptide and 56 amino acids from the carboxy-terminus of bovine wild-type PH20 during post-translational modification. BTH is also a glycoprotein and, based on its total composition comprising amino acids, has a mannose content of 5% and a glucosamine content of 2.2% (Borders and Raftery, 1968). When hyaluronidase derived from animals is repeatedly administered to the human body in high doses, neutralizing antibodies will be produced, and other biological materials derived from animals as impurities in addition to PH20 may cause allergic reactions. In particular, the use of PH20 extracted from cattle is limited due to concerns about mad cow disease. In order to overcome these problems, studies on recombinant human PH20 protein have been conducted.

It has been reported that recombinant human PH20 protein is expressed in yeast ( P. pastoris ), DS-2 insect cells, animal cells, etc. (Chen et al., 2016, Hofinger et al., 2007). Recombinant PH20 proteins produced by insect cells and yeast differ from human PH20 in their N-glycosidation patterns during post-translational modifications.

Among hyaluronidases, the protein structures of Hyal1 (PDB ID: 2PE4) (Chao et al., 2007) and bee venom hyaluronidase (PDB ID: 1FCQ, 1FCU, 1FCV) have been determined. Hyal1 consists of two structural domains, the catalytic domain and the EGF-like domain, and the catalytic domain is in the form of (β/α) ₈ (Chao et al., 2007), where the α-helix and β-strand represent the secondary structure of the protein. The EGF-like domain was fully conserved in variants in which the carboxy-terminus of Hyal1 was truncated differently. The amino acid sequences of Hyal1 and PH20 are 35.1% identical, and the protein tertiary structure of PH20 has not yet been discovered.

In the study of the structure/function relationship of human PH20, it was found that the carboxy-terminal domain of PH20 is very important for protein expression and enzyme activity. In particular, it has been reported that the carboxy-terminal end with amino acids 477-483 is very important for enzyme expression and activity ( Frost, 2007). The activity of full-length PH20 (amino acids 1-509) or a PH20 variant carboxy-terminally truncated at position 467 was only 10% of the activity of a PH20 variant carboxy-terminally truncated at one of positions 477 to 483 or lower (Frost, 2007). Halozyme Therapeutics has developed rHuPH20 (amino acids 36-482), a recombinant protein in which mature PH20 is cleaved at the carboxy terminus of Y482 (Bookbinder et al., 2006; Frost, 2007).

Meanwhile, the low stability of human PH20 itself remains unsolved, although research is ongoing to develop various therapeutic drugs in the form of subcutaneous injections using human PH20.

Against this technical background, the inventors of the present invention confirmed that human PH20 variants comprising one or more amino acid residue substitutions have very high enzymatic activity and thermal stability, and therefore filed a patent application (PCT/KR 2019 /009215), wherein one or more amino acid residues are substituted in the amino acid sequence of wild-type hyaluronidase PH20 in the α-helix 8 domain (S347 to C381) and the link between α-helix 7 and α-helix 8 domain (A333 to R346), and some amino acids at the amino-terminal and/or carboxy-terminal of PH20 are cleaved in human PH20 variants.

The inventors of the present invention also confirmed that the PH20 variants according to the present invention can be applied to pharmaceutical compositions or preparations containing drugs, such as antibody drugs, especially high doses of anti-HER2 antibodies or immune checkpoint antibodies. Therefore, according to the present invention, pharmaceutical compositions and preparations comprising PH20 variants and drugs such as anti-HER2 antibodies or immune checkpoint antibodies can be used for subcutaneous injections. The activities of drugs such as antibody drugs and PH20 variants are very stable and can be Maintain for a long time, and then complete the present invention.

The present invention is accomplished in view of the aforementioned problems. One object of the present invention is to provide a novel pharmaceutical composition, especially a pharmaceutical composition that can be used for subcutaneous injection. The pharmaceutical composition includes a PH20 variant with improved enzyme activity and thermal stability And drugs, wherein the thermal stability and activity of drugs and PH20 variants can be maintained for a long time.

Another object of the present invention is to provide a method for treating diseases, comprising administering the pharmaceutical composition according to the present invention to a subject in need of treatment.

According to the present invention, the above and other objects can be achieved by a pharmaceutical composition comprising (a) a drug and (b) a PH20 variant.

The PH20 variant contained in the pharmaceutical composition according to the present invention comprises a wild type human PH20 having the amino acid sequence of SEQ ID NO: 1 selected from the group consisting of S343E, M345T, K349E, L353A, L354I, N356E and I361T One or more amino acid residues of the group are substituted, and in one of the domains selected from α-helix 8 (S347 to C381) and/or the linking domain between α-helix 7 and α-helix 8 (A333 to R346) One or more regions may further include amino acid residue substitution, wherein some amino acid residues at the amino terminus and/or carboxyl terminus are selectively cleaved.

The pharmaceutical composition according to the present invention may further comprise one or more selected from pharmaceutically acceptable additives, especially buffers, stabilizers and surfactants.

The pharmaceutical composition according to the present invention can be used in the form of injection preparations for subcutaneous injection.

Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. Generally, the nomenclature used herein is that which is known and commonly used in the art.

One embodiment of the present invention relates to a pharmaceutical composition, including (a) a drug and (b) a PH20 variant. The pharmaceutical composition according to the present invention can be used to prevent or treat diseases, and is preferably suitable for subcutaneous injection.

The PH20 variant contained in the pharmaceutical composition according to the present invention is in (with the amino acid sequence of SEQ ID NO: 1) in the amino acid sequence of wild-type PH20, preferably in (with the amino acid sequence represented by SEQ ID NO: 1 In the sequence consisting of L36 to S490 in the amino acid sequence) in the amino acid sequence of the mature wild-type PH20, there are some amino acid residue substitutions in the region corresponding to the α-helical domain and/or its connecting domain, Preferably in the α-helix 8 domain (S347 to C381) and/or the linking domain (A333 to R346) between α-helix 7 and α-helix 8, more preferably in the amino acid region among T341 to N363 , optimally at T341 to I361, L342 to I361, S343 to I361, I344 to I361, M345 to I361 or M345 to N363.

In the present invention, "mature wild-type PH20" refers to a protein comprising amino acid residues L36 to S490 of SEQ ID NO: 1, which lacks the amine in wild-type PH20 having the sequence of SEQ ID NO: 1 In the amino acid sequence, M1 to T35 that form the message peptide and A491 to L509 that have nothing to do with the actual function of PH20.

Table 1, the amino acid sequence of wild-type PH20 (SEQ ID NO: 1) MGVLKFKHIFFRSFVKSSGVSQIVFTFLLIPCCLTLNFRAPPVIPNVPFLWAWNAPSEFCLGKFDEPLDMSLFSFIGSPRINATGQGVTIFYVDRLGYYPYIDSITGVTVNGGIPQKISLQDHLDKAKKDITFYMPVDNLGMAVIDWEEWRPTWARNWKPKDVYKNRSIELVQQQNVQLSLTEATEKAKQEFEKAGKDFLVETIKLGKLLRPNHLWGYYLFPDCYNHHYKKPGYNGSCFNVEIKRNDDLSWLWNESTALYPSIYLNTQQSPVAATLYVRNRVREAIRVSKIPDAKSPLPVFAYTRIVFTDQVLKFLSQDELVYTFGETVALGASGIVIWGTLSIMRSMKSCLLLDNYMETILNPYIINVTLAAKMCSQVLCQEQGVCIRKNWNSSDYLHLNPDNFAIQLEKGGKFTVRGKPTLEDLEQFSEKFYCSCYSTLSCKEKADVKDTDAVDVCIADGVCIDAFLKPPMETEEPQIFYNASPSTLSATMFIVSILFLIISSVASL

Specifically, the PH20 variants and fragments thereof contained in the pharmaceutical composition according to the present invention contain one or more mutations in the wild-type PH20 having the sequence of SEQ ID NO: 1, preferably selected from S343E, M345T, The substitution of amino acid residues of the group consisting of K349E, L353A, L354I, N356E and I361T is optimally substituted by one or more amino acid residues selected from the group consisting of L354I and N356E.

In the present invention, the term "PH20 variant" is intended to include mutations of some amino acid residues, preferably substitutions of amino acid residues in the sequence of wild-type human PH20, and The substitution of residues together with the deletion of some amino acid residues at the amino-terminus and/or carboxyl-terminus are used in substantially the same sense as the expression "PH20 variant or fragment thereof".

The inventors of the present invention have confirmed that, according to the experimental results, when the amino acid sequence part of the α-helix 8 domain of human PH20 and the linking domain between α-helix 7 and α-helix 8 is replaced by Hyal1 with high hydrophilicity When the amino acid sequence of the α-helix 8 domain and the connecting domain between α-helix 7 and α-helix 8 is substituted, the enzyme activity and protein aggregation temperature (protein aggregation temperature, Tagg) will increase at neutral pH, through which Previous studies may provide novel PH20 variants and fragments thereof that have improved enzymatic activity and thermostability compared to wild-type PH20.

Therefore, the PH20 variant contained in the pharmaceutical composition according to the present invention is in (with the amino acid sequence of SEQ ID NO: 1) in the amino acid sequence of wild-type PH20, preferably in (with the amino acid sequence represented by SEQ ID NO : the sequence consisting of L36 to S490 in the amino acid sequence of 1) in the amino acid sequence of mature wild-type PH20, comprising a group selected from the group consisting of S343E, M345T, K349E, L353A, L354I, N356E and I361T One or more amino acid residue substitutions, preferably one or more amino acid residue substitutions selected from the group consisting of L354I and N356E.

Among them, in the region corresponding to the α-helix domain and/or its connecting domain, preferably in the α-helix 8 domain (S347 to C381) and/or the connecting domain between α-helix 7 and α-helix 8 (A333 to R346), more preferably in the amino acid region corresponding to T341 to N363, T341 to I361, L342 to I361, S343 to I361, I344 to I361, M345 to I361 or M345 to N363, one or more amino acid residues base is substituted.

Specifically, in the PH20 variant comprised in the pharmaceutical composition according to the present invention, in wild-type PH20, preferably in mature wild-type PH20, the α-helix 8 domain (S347 to C381) and/or the mediator The linking domain (A333 to R346) between α-helix 7 and α-helix 8 can be substituted by some amino acid residues of Hyal1 having the sequence of SEQ ID NO: 51 (see Table 2 and Table 3) Some amino acid residues in the amino acid sequence of the corresponding region, but the present invention is not limited thereto.

Table 2, the amino acid sequence of wild type Hyal1 (SEQ ID NO: 51) MAAHLLPICALFLTLLDMAQGFRGPLLPNRPFTTVWNANTQWCLERHGVDVDVSVFDVVANPGQTFRGPDMTIFYSSQLGTYPYYTPTGEPVFGGLPQNASLIAHLARTFQDILAAIPAPDFSGLAVIDWEAWRPRWAFNWDTKDIYRQRSRALVQAQHPDWPAPQVEAVAQDQFQGAARAWMAGTLQLGRALRPRGLWGFYGFPDCYNYDFLSPNYTGQCPSGIRAQNDQLGWLWGQSRALYPSIYMPAVLEGTGKSQMYVQHRVAEAFRVAVAAGDPNLPVLPYVQIFYDTTNHFLPLDELEHSLGESAAQGAAGVVLWVSWENTRTKESCQAIKEYMDTTLGPFILNVTSGALLCSQALCSGHGRCVRRTSHPKALLLLNPASFSIQLTPGGGPLSLRGALSLEDQAQMAVEFKCRCYPGWQAPWCERKSMW

Table 3, Comparison between the amino acid sequences of PH20 and Hyal1 alpha helix Amino acid sequence of PH20 Amino acid sequence of Hyal1 alpha helix 1 P56 to D65 N39 to G48 alpha helix 3 S119 to M135 S101 to I117 alpha helix 4' K161 to N176 K144 to H159 alpha helix 4 S180 to R211 P163 to R194 alpha helix 5 F239 to S256 P222 to S239 alpha helix 6 A274 to D293 K257 to G277 alpha helix 7 S317 to G332 P299 to G314 alpha helix 8 S347 to C381 T329 to C363

More specifically, the PH20 variant or fragment thereof contained in the pharmaceutical composition according to the present invention is in the amino acid sequence of wild-type PH20, preferably in the amino acid sequence of mature wild-type PH20, preferably It preferably includes substitution of amino acid residues of L354I and/or N356E; and, preferably further comprises substitution of amino acid residues at one or more positions selected from T341 to N363, preferably at one or more positions selected from T341, One or more positions of the group consisting of L342, S343, I344, M345, S347, M348, K349, L352, L353, D355, E359, I361 and N363, but the present invention is not limited thereto; and, preferably further comprises Substitution of one or more amino acid residues selected from the group consisting of T341S, L342W, S343E, I344N, M345T, S347T, M348K, K349E, L352Q, L353A, D355K, E359D, I361T and N363G, but the present invention is not limited to this.

Preferably, the PH20 variant contained in the pharmaceutical composition according to the present invention may comprise amino acid residue substitutions selected from M345T, S347T, M348K, K349E, L352Q, L353A, L354I, D355K, N356E, E359D and I361T; Moreover, one or more amino acid residues selected from the group consisting of T341S, L342W, S343E, I344N and N363G may be further substituted, but the present invention is not limited thereto.

More preferably, the PH20 variant or fragment thereof contained in the pharmaceutical composition according to the present invention may include, but not limited to, any substitution selected from the following groups: (a) T341S, L342W, S343E, I344N, M345T, S347T, M348K, K349E, L352Q, L353A, L354I, D355K, N356E, E359D and I361T; (b) L342W, S343E, I344N, M345T, S347T, M348K, K349E, L352Q, L353A, L354I, D355K, N356E, E359D and I361T; (c) M345T, S347T, M348K, K349E, L352Q, L353A, L354I, D355K, N356E, E359D and I361T; (d) M345T, S347T, M348K, K349E, L352Q, L353A, L354I, D355K, N356E, E359D, I361T and N363G; (e) I344N, M345T, S347T, M348K, K349E, L352Q, L353A, L354I, D355K, N356E, E359D and I361T; and (f) S343E, I344N, M345T, S347T, M348K, K349E, L352Q, L353A, L354I, D355K, N356E, E359D and I361T.

In the present invention, the expression of the one-letter amino acid residue code together with the number, such as "S347", indicates the corresponding position of the amino acid residue in the amino acid sequence of SEQ ID NO: 1.

For example, "S347" means that the amino acid residue at position 347 in the amino acid sequence of SEQ ID NO: 1 is serine. In addition, "S347T" means that the serine at position 347 of SEQ ID NO: 1 is substituted with threonine.

The PH20 variant contained in the pharmaceutical composition according to the present invention is interpreted as a variant comprising a conservatively substituted amino acid residue at a specific amino acid residue position.

The term "conservative substitution" as used herein refers to a modification of a PH20 variant involving substitution of one or more amino acids with similar biochemical properties, which does not result in loss of the corresponding PH20 The biological or biochemical function of the variant.

A "retained amino acid substitution" is one in which an amino acid residue is replaced by an amino acid residue having a similar side chain. Families of amino acid residues having similar side chains have been defined and are known in the art. These families include amino acids with basic side chains (such as lysine, arginine, and histidine), amino acids with acidic side chains (such as aspartic acid and glutamic acid), amino acids with and without Amino acids with polar side chains (such as glycine, asparagine, glutamine, serine, threonine, tyrosine, and cysteine), amines with nonpolar side chains amino acids (such as alanine, valine, leucine, isoleucine, proline, phenylalanine, methionine, and tryptophan), amino acids with β-branched side chains (such as threo amine, valine, and isoleucine) and amino acids with aromatic side chains (such as tyrosine, phenylalanine, tryptophan, and histidine).

It is expected that the PH20 variants comprised in the pharmaceutical compositions according to the invention retain their activity despite the retained amino acid substitutions.

Furthermore, the PH20 variant or fragment thereof contained in the pharmaceutical composition according to the present invention is interpreted as comprising the PH20 variant or fragment thereof, which has substantially the same function or action as the PH20 variant or fragment thereof according to the present invention And/or functional, and have at least 80% or 85% amino acid sequence homology (amino acid sequence homology), preferably at least 90%, more preferably At least 95%, optimally at least 99%.

The PH20 variants according to the invention have an increased degree of expression in animal cells as well as an increased rate of protein folding and thus increased thermostability compared to mature wild-type PH20. Furthermore, despite the increased thermostability, the enzymatic activity of the PH20 variant exceeded or was similar to that of the wild-type PH20.

Meanwhile, it is known that when some amino acids, such as S490, at the carboxy terminus of mature wild-type PH20 are additionally excised, the enzyme activity will decrease, but the PH20 variant according to the present invention has When having an additional excised sequence at the end, PH20 still exhibits increased thermostability and increased or similar enzymatic activity compared to the mature wild-type PH20. In addition, the PH20 variant retained its enzymatic activity when up to five amino acid residues were excised from the amino-terminal amino acid, suggesting that residues from P41 on the amino-terminus play a role in protein expression and enzymatic activity. important role.

Therefore, the PH20 variant contained in the pharmaceutical composition according to the present invention is in the α-helix 8 domain (S347 to C381) and/or the linking domain (A333 to R346) between α-helix 7 and α-helix 8 of wild-type PH20. ) contains substitutions of some amino acid residues and deletion of some amino acid residues at the carboxy terminus and/or amino terminus, but the present invention is not limited thereto.

In some embodiments, the PH20 variant included in the pharmaceutical composition according to the present invention may comprise deletion of some amino acid residues at the amino terminus and/or deletion of some amino acid residues at the carboxy terminus, wherein some amino acid residues at the amino terminus The deletion of amino acid residues is derived from the cleavage (cleavage) before the amino acid residues selected from the group consisting of M1 to P42 at the amino terminus of the amino acid sequence of SEQ ID NO: 1, preferably Before amino acid residues L36, N37, F38, R39, A40, P41 or P42, while some amino acid deletions at the carboxy-terminus are derived from amino acids selected from the group consisting of V455 to W509 at the carboxyl-terminus Excision after residues, preferably after residues selected from the group consisting of V455 to S490, optimally after V455, C458, D461, C464, I465, D466, A467, F468, K470, P471, P472, M473, E474 , T475, E476, P478, I480, Y482, A484, P486, T488 or S490.

The expression "excision before L36, N37, F38, R39, A40, P41 or P42 at the amino terminus" means all amines from M1 to T35 immediately before L36 in the amino acid sequence of SEQ ID NO: 1, respectively all amino acid residues from M1 to L36 immediately before N37, all amino acid residues from M1 to N37 immediately before F38, all amino acid residues from M1 to F38 immediately before R39 residues, all amino acid residues from M1 to R39 immediately before A40, all amino acid residues from M1 to A40 immediately before P41, or all amino acid residues from M1 to P41 immediately before P42 was cut off and removed. The expression "cleavage before M1 at the amino-terminus of SEQ ID NO: 1" means that no cleavage occurs at the amino-terminus.

In addition, "V455, C458, D461, C464, I465, D466, A467, F468, K470, P471, P472, M473, E474, T475, E476, P478, I480, Y482, A484, P486, T488 or S490 at the carboxy-terminal The expression "excision after" means to cut off and remove V455, C458, D461, C464, I465, D466, A467, F468, K470, P472, M473, E474, T475, E476, Amino acid residues after P478, I480, Y482, A484, P486, T488 or S490. For example, a cut at S490 means a cut between S490 and A491.

Preferably, the human PH20 variant comprised in the pharmaceutical composition according to the present invention may have an amino acid sequence selected from the group consisting of the amino acid sequences of SEQ ID NOS: 5 to 50, preferably having SEQ ID NOS: NO: Amino acid sequence of 44, but the present invention is not limited thereto. In the PH20 variants constructed according to specific embodiments of the present invention, the sequences of substituted or excised amino acids are shown in Table 4 below.

Table 4, amino acid sequences and substitution/excision properties of PH20 variants according to the present invention name serial number replace sequence HM1 5 Twelve amino acids were substituted as M345T, S347T, M348K, K349E, L352Q, L353A, L354I, D355K, N356E, E359D, I361T and N363G. LNFRAPPVIPNVPFLWAWNAPSEFCLGKFDEPLDMSLFSFIGSPRINATGQGVTIFYVDRLGYYPYIDSITGVTVNGGIPQKISLQDHLDKAKKDITFYMPVDNLGMAVIDWEEWRPTWARNWKPKDVYKNRSIELVQQQNVQLSLTEATEKAKQEFEKAGKDFLVETIKLGKLLRPNHLWGYYLFPDCYNHHYKKPGYNGSCFNVEIKRNDDLSWLWNESTALYPSIYLNTQQSPVAATLYVRNRVREAIRVSKIPDAKSPLPVFAYTRIVFTDQVLKFLSQDELVYTFGETVALGASGIVIWGTLSI T R TKE SC QAIKE YM D T T L G PYIINVTLAAKMCSQVLCQEQGVCIRKNWNSSDYLHLNPDNFAIQLEKGGKFTVRGKPTLEDLEQFSEKFYCSCYSTLSCKEKADVKDTDAVDVCIADGVCIDAFLKPPMETEEPQIFYNASPSTLS HM2 6 Seven amino acids were substituted as Y365F, I367L, L371S, A372G, K374L, M375L and V379A. LNFRAPPVIPNVPFLWAWNAPSEFCLGKFDEPLDMSLFSFIGSPRINATGQGVTIFYVDRLGYYPYIDSITGVTVNGGIPQKISLQDHLDKAKKDITFYMPVDNLGMAVIDWEEWRPTWARNWKPKDVYKNRSIELVQQQNVQLSLTEATEKAKQEFEKAGKDFLVETIKLGKLLRPNHLWGYYLFPDCYNHHYKKPGYNGSCFNVEIKRNDDLSWLWNESTALYPSIYLNTQQSPVAATLYVRNRVREAIRVSKIPDAKSPLPVFAYTRIVFTDQVLKFLSQDELVYTFGETVALGASGIVIWGTLSITRTKESCQAIKEYMDTTLNP F I L NVT SG A LL CSQ A LCQEQGVCIRKNWNSSDYLHLNPDNFAIQLEKGGKFTVRGKPTLEDLEQFSEKFYCSCYSTLSCKEKADVKDTDAVDVCIADGVCIDAFLKPPMETEEPQIFYNASPSTLS HM3 7 Nineteen amino acids were substituted as M345T, S347T, M348K, K349E, L352Q, L353A, L354I, D355K, N356E, E359D, I361T, N363G, Y365F, I367L, L371S, A372G, K374L, M375L and V379A. LNFRAPPVIPNVPFLWAWNAPSEFCLGKFDEPLDMSLFSFIGSPRINATGQGVTIFYVDRLGYYPYIDSITGVTVNGGIPQKISLQDHLDKAKKDITFYMPVDNLGMAVIDWEEWRPTWARNWKPKDVYKNRSIELVQQQNVQLSLTEATEKAKQEFEKAGKDFLVETIKLGKLLRPNHLWGYYLFPDCYNHHYKKPGYNGSCFNVEIKRNDDLSWLWNESTALYPSIYLNTQQSPVAATLYVRNRVREAIRVSKIPDAKSPLPVFAYTRIVFTDQVLKFLSQDELVYTFGETVALGASGIVIWGTLSI T R TKE SC QAIKE YM D T T L G P F I L NVT SG A LL CSQ A LCQEQGVCIRKNWNSSDYLHLNPDNFAIQLEKGGKFTVRGKPTLEDLEQFSEKFYCSCYSTLSCKEKADVKDTDAVDVCIADGVCIDAFLKPPMETEEPQIFYNASPSTLS HM4 8 Seventeen amino acids were substituted as G340V, T341S, L342W, S343E, I344N, M345T, S347T, M348K, K349E, L352Q, L353A, L354I, D355K, N356E, E359D, I361T and N363G. LNFRAPPVIPNVPFLWAWNAPSEFCLGKFDEPLDMSLFSFIGSPRINATGQGVTIFYVDRLGYYPYIDSITGVTVNGGIPQKISLQDHLDKAKKDITFYMPVDNLGMAVIDWEEWRPTWARNWKPKDVYKNRSIELVQQQNVQLSLTEATEKAKQEFEKAGKDFLVETIKLGKLLRPNHLWGYYLFPDCYNHHYKKPGYNGSCFNVEIKRNDDLSWLWNESTALYPSIYLNTQQSPVAATLYVRNRVREAIRVSKIPDAKSPLPVFAYTRIVFTDQVLKFLSQDELVYTFGETVALGASGIVIW VSWENT R TKE SC QAIKE YM D T T L G PYIINVTLAAKMCSQVLCQEQGVCIRKNWNSSDYLHLNPDNFAIQLEKGGKFTVRGKPTLEDLEQFSEKFYCSCYSTLSCKEKADVKDTDAVDVCIADGVCIDAFLKPPMETEEPQIFYNASPSTLS HM6 9 Eleven amino acid residues were substituted as M345T, S347T, M348K, K349E, L352Q, L353A, L354I, D355K, N356E, E359D and I361T. LNFRAPPVIPNVPFLWAWNAPSEFCLGKFDEPLDMSLFSFIGSPRINATGQGVTIFYVDRLGYYPYIDSITGVTVNGGIPQKISLQDHLDKAKKDITFYMPVDNLGMAVIDWEEWRPTWARNWKPKDVYKNRSIELVQQQNVQLSLTEATEKAKQEFEKAGKDFLVETIKLGKLLRPNHLWGYYLFPDCYNHHYKKPGYNGSCFNVEIKRNDDLSWLWNESTALYPSIYLNTQQSPVAATLYVRNRVREAIRVSKIPDAKSPLPVFAYTRIVFTDQVLKFLSQDELVYTFGETVALGASGIVIWGTLSI T R TKE SC QAIKE YM D T T LNPYIINVTLAAKMCSQVLCQEQGVCIRKNWNSSDYLHLNPDNFAIQLEKGGKFTVRGKPTLEDLEQFSEKFYCSCYSTLSCKEKADVKDTDAVDVCIADGVCIDAFLKPPMETEEPQIFYNASPSTLS HM7 10 Sixteen amino acids were substituted as G340V, T341S, L342W, S343E, I344N, M345T, S347T, M348K, K349E, L352Q, L353A, L354I, D355K, N356E, E359D and I361T. LNFRAPPVIPNVPFLWAWNAPSEFCLGKFDEPLDMSLFSFIGSPRINATGQGVTIFYVDRLGYYPYIDSITGVTVNGGIPQKISLQDHLDKAKKDITFYMPVDNLGMAVIDWEEWRPTWARNWKPKDVYKNRSIELVQQQNVQLSLTEATEKAKQEFEKAGKDFLVETIKLGKLLRPNHLWGYYLFPDCYNHHYKKPGYNGSCFNVEIKRNDDLSWLWNESTALYPSIYLNTQQSPVAATLYVRNRVREAIRVSKIPDAKSPLPVFAYTRIVFTDQVLKFLSQDELVYTFGETVALGASGIVIW VSWENT R TKE SC QAIKE YM D T T LNPYIINVTLAAKMCSQVLCQEQGVCIRKNWNSSDYLHLNPDNFAIQLEKGGKFTVRGKPTLEDLEQFSEKFYCSCYSTLSCKEKADVKDTDAVDVCIADGVCIDAFLKPPMETEEPQIFYNASPSTLS HM8 11 Twelve amino acids were substituted as I344N, M345T, S347T, M348K, K349E, L352Q, L353A, L354I, D355K, N356E, E359D and I361T. LNFRAPPVIPNVPFLWAWNAPSEFCLGKFDEPLDMSLFSFIGSPRINATGQGVTIFYVDRLGYYPYIDSITGVTVNGGIPQKISLQDHLDKAKKDITFYMPVDNLGMAVIDWEEWRPTWARNWKPKDVYKNRSIELVQQQNVQLSLTEATEKAKQEFEKAGKDFLVETIKLGKLLRPNHLWGYYLFPDCYNHHYKKPGYNGSCFNVEIKRNDDLSWLWNESTALYPSIYLNTQQSPVAATLYVRNRVREAIRVSKIPDAKSPLPVFAYTRIVFTDQVLKFLSQDELVYTFGETVALGASGIVIWGTLS NT R TKE SC QAIKE YM D T T LNPYIINVTLAAKMCSQVLCQEQGVCIRKNWNSSDYLHLNPDNFAIQLEKGGKFTVRGKPTLEDLEQFSEKFYCSCYSTLSCKEKADVKDTDAVDVCIADGVCIDAFLKPPMETEEPQIFYNASPSTLS HM9 12 Thirteen amino acids were substituted as S343E, I344N, M345T, S347T, M348K, K349E, L352Q, L353A, L354I, D355K, N356E, E359D and I361T. LNFRAPPVIPNVPFLWAWNAPSEFCLGKFDEPLDMSLFSFIGSPRINATGQGVTIFYVDRLGYYPYIDSITGVTVNGGIPQKISLQDHLDKAKKDITFYMPVDNLGMAVIDWEEWRPTWARNWKPKDVYKNRSIELVQQQNVQLSLTEATEKAKQEFEKAGKDFLVETIKLGKLLRPNHLWGYYLFPDCYNHHYKKPGYNGSCFNVEIKRNDDLSWLWNESTALYPSIYLNTQQSPVAATLYVRNRVREAIRVSKIPDAKSPLPVFAYTRIVFTDQVLKFLSQDELVYTFGETVALGASGIVIWGTL ENT R TKE SC QAIKE YM D T T LNPYIINVTLAAKMCSQVLCQEQGVCIRKNWNSSDYLHLNPDNFAIQLEKGGKFTVRGKPTLEDLEQFSEKFYCSCYSTLSCKEKADVKDTDAVDVCIADGVCIDAFLKPPMETEEPQIFYNASPSTLS HMl0 13 Fourteen amino acid residues were substituted as L342W, S343E, I344N, M345T, S347T, M348K, K349E, L352Q, L353A, L354I, D355K, N356E, E359D and I361T. LNFRAPPVIPNVPFLWAWNAPSEFCLGKFDEPLDMSLFSFIGSPRINATGQGVTIFYVDRLGYYPYIDSITGVTVNGGIPQKISLQDHLDKAKKDITFYMPVDNLGMAVIDWEEWRPTWARNWKPKDVYKNRSIELVQQQNVQLSLTEATEKAKQEFEKAGKDFLVETIKLGKLLRPNHLWGYYLFPDCYNHHYKKPGYNGSCFNVEIKRNDDLSWLWNESTALYPSIYLNTQQSPVAATLYVRNRVREAIRVSKIPDAKSPLPVFAYTRIVFTDQVLKFLSQDELVYTFGETVALGASGIVIWGT WENT R TKE SC QAIKE YM D T T LNPYIINVTLAAKMCSQVLCQEQGVCIRKNWNSSDYLHLNPDNFAIQLEKGGKFTVRGKPTLEDLEQFSEKFYCSCYSTLSCKEKADVKDTDAVDVCIADGVCIDAFLKPPMETEEPQIFYNASPSTLS HM11 14 Thirteen amino acid residues were substituted as M345T, S347T, M348K, K349E, L352Q, L353A, L354I, D355K, N356E, E359D, I361T, Y365F and I367L. LNFRAPPVIPNVPFLWAWNAPSEFCLGKFDEPLDMSLFSFIGSPRINATGQGVTIFYVDRLGYYPYIDSITGVTVNGGIPQKISLQDHLDKAKKDITFYMPVDNLGMAVIDWEEWRPTWARNWKPKDVYKNRSIELVQQQNVQLSLTEATEKAKQEFEKAGKDFLVETIKLGKLLRPNHLWGYYLFPDCYNHHYKKPGYNGSCFNVEIKRNDDLSWLWNESTALYPSIYLNTQQSPVAATLYVRNRVREAIRVSKIPDAKSPLPVFAYTRIVFTDQVLKFLSQDELVYTFGETVALGASGIVIWGTLSI T R TKE SC QAIKE YM D T T LNP F I L NVTLAAKMCSQVLCQEQGVCIRKNWNSSDYLHLNPDNFAIQLEKGGKFTVRGKPTLEDLEQFSEKFYCSCYSTLSCKEKADVKDTDAVDVCIADGVCIDAFLKPPMETEEPQIFYNASPSTLS HM12 15 Fifteen amino acid residues were substituted as M345T, S347T, M348K, K349E, L352Q, L353A, L354I, D355K, N356E, E359D, I361T, Y365F, I367L, L371S and A372G. LNFRAPPVIPNVPFLWAWNAPSEFCLGKFDEPLDMSLFSFIGSPRINATGQGVTIFYVDRLGYYPYIDSITGVTVNGGIPQKISLQDHLDKAKKDITFYMPVDNLGMAVIDWEEWRPTWARNWKPKDVYKNRSIELVQQQNVQLSLTEATEKAKQEFEKAGKDFLVETIKLGKLLRPNHLWGYYLFPDCYNHHYKKPGYNGSCFNVEIKRNDDLSWLWNESTALYPSIYLNTQQSPVAATLYVRNRVREAIRVSKIPDAKSPLPVFAYTRIVFTDQVLKFLSQDELVYTFGETVALGASGIVIWGTLSI T R TKE SC QAIKE YM D T T LNP F I L NVT SG AKMCSQVLCQEQGVCIRKNWNSSDYLHLNPDNFAIQLEKGGKFTVRGKPTLEDLEQFSEKFYCSCYSTLSCKEKADVKDTDAVDVCIADGVCIDAFLKPPMETEEPQIFYNASPSTLS HM13 16 Eleven amino acid residues were substituted as M345T, S347T, M348K, K349E, L352Q, L353A, L354I, D355K, N356E, E359D, and I361T, and cleaved before the amino-terminal residue F38. FRAPPVIPNVPFLWAWNAPSEFCLGKFDEPLDMSLFSFIGSPRINATGQGVTIFYVDRLGYYPYIDSITGVTVNGGIPQKISLQDHLDKAKKDITFYMPVDNLGMAVIDWEEWRPTWARNWKPKDVYKNRSIELVQQQNVQLSLTEATEKAKQEFEKAGKDFLVETIKLGKLLRPNHLWGYYLFPDCYNHHYKKPGYNGSCFNVEIKRNDDLSWLWNESTALYPSIYLNTQQSPVAATLYVRNRVREAIRVSKIPDAKSPLPVFAYTRIVFTDQVLKFLSQDELVYTFGETVALGASGIVIWGTLSI T R TKE SC QAIKE YM D T T LNPYIINVTLAAKMCSQVLCQEQGVCIRKNWNSSDYLHLNPDNFAIQLEKGGKFTVRGKPTLEDLEQFSEKFYCSCYSTLSCKEKADVKDTDAVDVCIADGVCIDAFLKPPMETEEPQIFYNASPSTLS HM14 17 Eleven amino acids were substituted into M345T, S347T, M348K, K349E, L352Q, L353A, L354I, D355K, N356E, E359D and I361T, and were cleaved after the carboxyl group of I465. LNFRAPPVIPNVPFLWAWNAPSEFCLGKFDEPLDMSLFSFIGSPRINATGQGVTIFYVDRLGYYPYIDSITGVTVNGGIPQKISLQDHLDKAKKDITFYMPVDNLGMAVIDWEEWRPTWARNWKPKDVYKNRSIELVQQQNVQLSLTEATEKAKQEFEKAGKDFLVETIKLGKLLRPNHLWGYYLFPDCYNHHYKKPGYNGSCFNVEIKRNDDLSWLWNESTALYPSIYLNTQQSPVAATLYVRNRVREAIRVSKIPDAKSPLPVFAYTRIVFTDQVLKFLSQDELVYTFGETVALGASGIVIWGTLSI T R TKE SC QAIKE YM D T T LNPYIINVTLAAKMCSQVLCQEQGVCIRKNWNSSDYLHLNPDNFAIQLEKGGKFTVRGKPTLEDLEQFSEKFYCSCYSTLSCKEKADVKDTDAVDVCIADGVCI HM15 18 Eleven amino acids were substituted into M345T, S347T, M348K, K349E, L352Q, L353A, L354I, D355K, N356E, E359D and I361T, and were cleaved after the carboxyl group of F468. LNFRAPPVIPNVPFLWAWNAPSEFCLGKFDEPLDMSLFSFIGSPRINATGQGVTIFYVDRLGYYPYIDSITGVTVNGGIPQKISLQDHLDKAKKDITFYMPVDNLGMAVIDWEEWRPTWARNWKPKDVYKNRSIELVQQQNVQLSLTEATEKAKQEFEKAGKDFLVETIKLGKLLRPNHLWGYYLFPDCYNHHYKKPGYNGSCFNVEIKRNDDLSWLWNESTALYPSIYLNTQQSPVAATLYVRNRVREAIRVSKIPDAKSPLPVFAYTRIVFTDQVLKFLSQDELVYTFGETVALGASGIVIWGTLSI T R TKE SC QAIKE YM D T T LNPYIINVTLAAKMCSQVLCQEQGVCIRKNWNSSDYLHLNPDNFAIQLEKGGKFTVRGKPTLEDLEQFSEKFYCSCYSTLSCKEKADVKDTDAVDVCIADGVCIDAF HM16 19 Eleven amino acids were substituted into M345T, S347T, M348K, K349E, L352Q, L353A, L354I, D355K, N356E, E359D and I361T, and were cleaved after the carboxyl group of P471. LNFRAPPVIPNVPFLWAWNAPSEFCLGKFDEPLDMSLFSFIGSPRINATGQGVTIFYVDRLGYYPYIDSITGVTVNGGIPQKISLQDHLDKAKKDITFYMPVDNLGMAVIDWEEWRPTWARNWKPKDVYKNRSIELVQQQNVQLSLTEATEKAKQEFEKAGKDFLVETIKLGKLLRPNHLWGYYLFPDCYNHHYKKPGYNGSCFNVEIKRNDDLSWLWNESTALYPSIYLNTQQSPVAATLYVRNRVREAIRVSKIPDAKSPLPVFAYTRIVFTDQVLKFLSQDELVYTFGETVALGASGIVIWGTLSI T R TKE SC QAIKE YM D T T LNPYIINVTLAAKMCSQVLCQEQGVCIRKNWNSSDYLHLNPDNFAIQLEKGGKFTVRGKPTLEDLEQFSEKFYCSCYSTLSCKEKADVKDTDAVDVCIADGVCIDAFLKP HM17 20 Amino acids L36 to V47 were substituted with FRGPLLPNR, and 11 amino acids were substituted with M345T, S347T, M348K, K349E, L352Q, L353A, L354I, D355K, N356E, E359D, and I361T. FRGPLLPNR PFLWAWNAPSEFCLGKFDEPLDMSLFSFIGSPRINATGQGVTIFYVDRLGYYPYIDSITGVTVNGGIPQKISLQDHLDKAKKDITFYMPVDNLGMAVIDWEEWRPTWARNWKPKDVYKNRSIELVQQQNVQLSLTEATEKAKQEFEKAGKDFLVETIKLGKLLRPNHLWGYYLFPDCYNHHYKKPGYNGSCFNVEIKRNDDLSWLWNESTALYPSIYLNTQQSPVAATLYVRNRVREAIRVSKIPDAKSPLPVFAYTRIVFTDQVLKFLSQDELVYTFGETVALGASGIVIWGTLSI T R TKE SC QAIKE YM D T T LNPYIINVTLAAKMCSQVLCQEQGVCIRKNWNSSDYLHLNPDNFAIQLEKGGKFTVRGKPTLEDLEQFSEKFYCSCYSTLSCKEKADVKDTDAVDVCIADGVCIDAFLKPPMETEEPQIFYNASPSTLS HM18 twenty one Amino acids L36 to A52 were substituted with FRGPLLPNRPFTTV and 11 amino acids were substituted with M345T, S347T, M348K, K349E, L352Q, L353A, L354I, D355K, N356E, E359D and I361T. FRGPLLPNRPFTTV WNAPSEFCLGKFDEPLDMSLFSFIGSPRINATGQGVTIFYVDRLGYYPYIDSITGVTVNGGIPQKISLQDHLDKAKKDITFYMPVDNLGMAVIDWEEWRPTWARNWKPKDVYKNRSIELVQQQNVQLSLTEATEKAKQEFEKAGKDFLVETIKLGKLLRPNHLWGYYLFPDCYNHHYKKPGYNGSCFNVEIKRNDDLSWLWNESTALYPSIYLNTQQSPVAATLYVRNRVREAIRVSKIPDAKSPLPVFAYTRIVFTDQVLKFLSQDELVYTFGETVALGASGIVIWGTLSI T R TKE SC QAIKE YM D T T LNPYIINVTLAAKMCSQVLCQEQGVCIRKNWNSSDYLHLNPDNFAIQLEKGGKFTVRGKPTLEDLEQFSEKFYCSCYSTLSCKEKADVKDTDAVDVCIADGVCIDAFLKPPMETEEPQIFYNASPSTLS HM19 twenty two Fourteen amino acid residues were substituted as L342W, S343E, I344N, M345T, S347T, M348K, K349E, L352Q, L353A, L354I, D355K, N356E, E359D and I361T, and before the amino-terminal residue F38 and at The carboxy-terminal residue K470 was followed by cleavage. FRAPPVIPNVPFLWAWNAPSEFCLGKFDEPLDMSLFSFIGSPRINATGQGVTIFYVDRLGYYPYIDSITGVTVNGGIPQKISLQDHLDKAKKDITFYMPVDNLGMAVIDWEEWRPTWARNWKPKDVYKNRSIELVQQQNVQLSLTEATEKAKQEFEKAGKDFLVETIKLGKLLRPNHLWGYYLFPDCYNHHYKKPGYNGSCFNVEIKRNDDLSWLWNESTALYPSIYLNTQQSPVAATLYVRNRVREAIRVSKIPDAKSPLPVFAYTRIVFTDQVLKFLSQDELVYTFGETVALGASGIVIWGT WENT R TKE SC QAIKE YM D T T LNPYIINVTLAAKMCSQVLCQEQGVCIRKNWNSSDYLHLNPDNFAIQLEKGGKFTVRGKPTLEDLEQFSEKFYCSCYSTLSCKEKADVKDTDAVDVCIADGVCIDAFLK HM20 twenty three Fourteen amino acid residues were substituted as L342W, S343E, I344N, M345T, S347T, M348K, K349E, L352Q, L353A, L354I, D355K, N356E, E359D and I361T, and before the amino-terminal residue F38 and at The carboxy-terminal residue F468 was then cleaved. FRAPPVIPNVPFLWAWNAPSEFCLGKFDEPLDMSLFSFIGSPRINATGQGVTIFYVDRLGYYPYIDSITGVTVNGGIPQKISLQDHLDKAKKDITFYMPVDNLGMAVIDWEEWRPTWARNWKPKDVYKNRSIELVQQQNVQLSLTEATEKAKQEFEKAGKDFLVETIKLGKLLRPNHLWGYYLFPDCYNHHYKKPGYNGSCFNVEIKRNDDLSWLWNESTALYPSIYLNTQQSPVAATLYVRNRVREAIRVSKIPDAKSPLPVFAYTRIVFTDQVLKFLSQDELVYTFGETVALGASGIVIWGT WENT R TKE SC QAIKE YM D T T LNPYIINVTLAAKMCSQVLCQEQGVCIRKNWNSSDYLHLNPDNFAIQLEKGGKFTVRGKPTLEDLEQFSEKFYCSCYSTLSCKEKADVKDTDAVDVCIADGVCIDAF HM21 twenty four Fifteen amino acid residues were substituted as T341S, L342W, S343E, I344N, M345T, S347T, M348K, K349E, L352Q, L353A, L354I, D355K, N356E, E359D and I361T. LNFRAPPVIPNVPFLWAWNAPSEFCLGKFDEPLDMSLFSFIGSPRINATGQGVTIFYVDRLGYYPYIDSITGVTVNGGIPQKISLQDHLDKAKKDITFYMPVDNLGMAVIDWEEWRPTWARNWKPKDVYKNRSIELVQQQNVQLSLTEATEKAKQEFEKAGKDFLVETIKLGKLLRPNHLWGYYLFPDCYNHHYKKPGYNGSCFNVEIKRNDDLSWLWNESTALYPSIYLNTQQSPVAATLYVRNRVREAIRVSKIPDAKSPLPVFAYTRIVFTDQVLKFLSQDELVYTFGETVALGASGIVIWG SWENT R TKE SC QAIKE YM D T T LNPYIINVTLAAKMCSQVLCQEQGVCIRKNWNSSDYLHLNPDNFAIQLEKGGKFTVRGKPTLEDLEQFSEKFYCSCYSTLSCKEKADVKDTDAVDVCIADGVCIDAFLKPPMETEEPQIFYNASPSTLS HM24 25 Eleven amino acid residues were substituted as M345T, S347T, M348K, K349E, L352Q, L353A, L354I, D355K, N356E, E359D, and I361T, and were cleaved before the amino-terminal residue A40. APPVIPNVPFLWAWNAPSEFCLGKFDEPLDMSLFSFIGSPRINATGQGVTIFYVDRLGYYPYIDSITGVTVNGGIPQKISLQDHLDKAKKDITFYMPVDNLGMAVIDWEEWRPTWARNWKPKDVYKNRSIELVQQQNVQLSLTEATEKAKQEFEKAGKDFLVETIKLGKLLRPNHLWGYYLFPDCYNHHYKKPGYNGSCFNVEIKRNDDLSWLWNESTALYPSIYLNTQQSPVAATLYVRNRVREAIRVSKIPDAKSPLPVFAYTRIVFTDQVLKFLSQDELVYTFGETVALGASGIVIWGTLSI T R TKE SC QAIKE YM D T T LNPYIINVTLAAKMCSQVLCQEQGVCIRKNWNSSDYLHLNPDNFAIQLEKGGKFTVRGKPTLEDLEQFSEKFYCSCYSTLSCKEKADVKDTDAVDVCIADGVCIDAFLKPPMETEEPQIFYNASPSTLS HM25 26 Eleven amino acids were substituted as M345T, S347T, M348K, K349E, L352Q, L353A, L354I, D355K, N356E, E359D, and I361T, and were cleaved before the amino-terminal residue P42. PVIPNVPFLWAWNAPSEFCLGKFDEPLDMSLFSFIGSPRINATGQGVTIFYVDRLGYYPYIDSITGVTVNGGIPQKISLQDHLDKAKKDITFYMPVDNLGMAVIDWEEWRPTWARNWKPKDVYKNRSIELVQQQNVQLSLTEATEKAKQEFEKAGKDFLVETIKLGKLLRPNHLWGYYLFPDCYNHHYKKPGYNGSCFNVEIKRNDDLSWLWNESTALYPSIYLNTQQSPVAATLYVRNRVREAIRVSKIPDAKSPLPVFAYTRIVFTDQVLKFLSQDELVYTFGETVALGASGIVIWGTLSI T R TKE SC QAIKE YM D T T LNPYIINVTLAAKMCSQVLCQEQGVCIRKNWNSSDYLHLNPDNFAIQLEKGGKFTVRGKPTLEDLEQFSEKFYCSCYSTLSCKEKADVKDTDAVDVCIADGVCIDAFLKPPMETEEPQIFYNASPSTLS HM29 27 Fourteen amino acid residues were substituted as L342W, S343E, I344N, M345T, S347T, M348K, K349E, L352Q, L353A, L354I, D355K, N356E, E359D, and I361T, and before the amino-terminal residue L36 and at The carboxy-terminal residue A467 was followed by cleavage. LNFRAPPVIPNVPFLWAWNAPSEFCLGKFDEPLDMSLFSFIGSPRINATGQGVTIFYVDRLGYYPYIDSITGVTVNGGIPQKISLQDHLDKAKKDITFYMPVDNLGMAVIDWEEWRPTWARNWKPKDVYKNRSIELVQQQNVQLSLTEATEKAKQEFEKAGKDFLVETIKLGKLLRPNHLWGYYLFPDCYNHHYKKPGYNGSCFNVEIKRNDDLSWLWNESTALYPSIYLNTQQSPVAATLYVRNRVREAIRVSKIPDAKSPLPVFAYTRIVFTDQVLKFLSQDELVYTFGETVALGASGIVIWGT WENT R TKE SC QAIKE YM D T T LNPYIINVTLAAKMCSQVLCQEQGVCIRKNWNSSDYLHLNPDNFAIQLEKGGKFTVRGKPTLEDLEQFSEKFYCSCYSTLSCKEKADVKDTDAVDVCIADGVCIDA HM30 28 Fourteen amino acid residues were substituted as L342W, S343E, I344N, M345T, S347T, M348K, K349E, L352Q, L353A, L354I, D355K, N356E, E359D, and I361T, and before the amino-terminal residue L36 and at The carboxy-terminal residue C464 was then cleaved. LNFRAPPVIPNVPFLWAWNAPSEFCLGKFDEPLDMSLFSFIGSPRINATGQGVTIFYVDRLGYYPYIDSITGVTVNGGIPQKISLQDHLDKAKKDITFYMPVDNLGMAVIDWEEWRPTWARNWKPKDVYKNRSIELVQQQNVQLSLTEATEKAKQEFEKAGKDFLVETIKLGKLLRPNHLWGYYLFPDCYNHHYKKPGYNGSCFNVEIKRNDDLSWLWNESTALYPSIYLNTQQSPVAATLYVRNRVREAIRVSKIPDAKSPLPVFAYTRIVFTDQVLKFLSQDELVYTFGETVALGASGIVIWGT WENT R TKE SC QAIKE YM D T T LNPYIINVTLAAKMCSQVLCQEQGVCIRKNWNSSDYLHLNPDNFAIQLEKGGKFTVRGKPTLEDLEQFSEKFYCSCYSTLSCKEKADVKDTDAVDVCIADGVC HM31 29 Fourteen amino acid residues were substituted as L342W, S343E, I344N, M345T, S347T, M348K, K349E, L352Q, L353A, L354I, D355K, N356E, E359D and I361T, and before the amino-terminal residue L36 and at the The carboxy-terminal residue D461 was then cleaved. LNFRAPPVIPNVPFLWAWNAPSEFCLGKFDEPLDMSLFSFIGSPRINATGQGVTIFYVDRLGYYPYIDSITGVTVNGGIPQKISLQDHLDKAKKDITFYMPVDNLGMAVIDWEEWRPTWARNWKPKDVYKNRSIELVQQQNVQLSLTEATEKAKQEFEKAGKDFLVETIKLGKLLRPNHLWGYYLFPDCYNHHYKKPGYNGSCFNVEIKRNDDLSWLWNESTALYPSIYLNTQQSPVAATLYVRNRVREAIRVSKIPDAKSPLPVFAYTRIVFTDQVLKFLSQDELVYTFGETVALGASGIVIWGT WENT R TKE SC QAIKE YM D T T LNPYIINVTLAAKMCSQVLCQEQGVCIRKNWNSSDYLHLNPDNFAIQLEKGGKFTVRGKPTLEDLEQFSEKFYCSCYSTLSCKEKADVKDTDAVDVCIAD HM32 30 Fourteen amino acid residues were substituted as L342W, S343E, I344N, M345T, S347T, M348K, K349E, L352Q, L353A, L354I, D355K, N356E, E359D, and I361T, and the amino-terminal residue L36 was preceded by the The carboxy-terminal residue C458 was then cleaved. LNFRAPPVIPNVPFLWAWNAPSEFCLGKFDEPLDMSLFSFIGSPRINATGQGVTIFYVDRLGYYPYIDSITGVTVNGGIPQKISLQDHLDKAKKDITFYMPVDNLGMAVIDWEEWRPTWARNWKPKDVYKNRSIELVQQQNVQLSLTEATEKAKQEFEKAGKDFLVETIKLGKLLRPNHLWGYYLFPDCYNHHYKKPGYNGSCFNVEIKRNDDLSWLWNESTALYPSIYLNTQQSPVAATLYVRNRVREAIRVSKIPDAKSPLPVFAYTRIVFTDQVLKFLSQDELVYTFGETVALGASGIVIWGT WENT R TKE SC QAIKE YM D T T LNPYIINVTLAAKMCSQVLCQEQGVCIRKNWNSSDYLHLNPDNFAIQLEKGGKFTVRGKPTLEDLEQFSEKFYCSCYSTLSCKEKADVKDTDAVDVC HM33 31 Fourteen amino acid residues were substituted as L342W, S343E, I344N, M345T, S347T, M348K, K349E, L352Q, L353A, L354I, D355K, N356E, E359D, and I361T, and before the amino-terminal residue L36 and the carboxyl The terminal residue V455 was followed by cleavage. LNFRAPPVIPNVPFLWAWNAPSEFCLGKFDEPLDMSLFSFIGSPRINATGQGVTIFYVDRLGYYPYIDSITGVTVNGGIPQKISLQDHLDKAKKDITFYMPVDNLGMAVIDWEEWRPTWARNWKPKDVYKNRSIELVQQQNVQLSLTEATEKAKQEFEKAGKDFLVETIKLGKLLRPNHLWGYYLFPDCYNHHYKKPGYNGSCFNVEIKRNDDLSWLWNESTALYPSIYLNTQQSPVAATLYVRNRVREAIRVSKIPDAKSPLPVFAYTRIVFTDQVLKFLSQDELVYTFGETVALGASGIVIWGT WENT R TKE SC QAIKE YM D T T LNPYIINVTLAAKMCSQVLCQEQGVCIRKNWNSSDYLHLNPDNFAIQLEKGGKFTVRGKPTLEDLEQFSEKFYCSCYSTLSCKEKADVKDTDAV HP34 32 Fifteen amino acid residues were substituted as T341S, L342W, S343E, I344N, M345T, S347T, M348K, K349E, L352Q, L353A, L354I, D355K, N356E, E359D, and I361T, and before the amino-terminal residue F38 and cleavage after the carboxy-terminal residue K470. FRAPPVIPNVPFLWAWNAPSEFCLGKFDEPLDMSLFSFIGSPRINATGQGVTIFYVDRLGYYPYIDSITGVTVNGGIPQKISLQDHLDKAKKDITFYMPVDNLGMAVIDWEEWRPTWARNWKPKDVYKNRSIELVQQQNVQLSLTEATEKAKQEFEKAGKDFLVETIKLGKLLRPNHLWGYYLFPDCYNHHYKKPGYNGSCFNVEIKRNDDLSWLWNESTALYPSIYLNTQQSPVAATLYVRNRVREAIRVSKIPDAKSPLPVFAYTRIVFTDQVLKFLSQDELVYTFGETVALGASGIVIWG SWENT R TKE SC QAIKE YM D T T LNPYIINVTLAAKMCSQVLCQEQGVCIRKNWNSSDYLHLNPDNFAIQLEKGGKFTVRGKPTLEDLEQFSEKFYCSCYSTLSCKEKADVKDTDAVDVCIADGVCIDAFLK HM35 33 Fourteen amino acid residues were substituted as L342W, S343E, I344N, M345T, S347T, M348K, K349E, L352Q, L353A, L354I, D355K, N356E, E359D and I361T, and before the amino-terminal residue F38 and at The carboxy-terminal residue P472 was then cleaved. FRAPPVIPNVPFLWAWNAPSEFCLGKFDEPLDMSLFSFIGSPRINATGQGVTIFYVDRLGYYPYIDSITGVTVNGGIPQKISLQDHLDKAKKDITFYMPVDNLGMAVIDWEEWRPTWARNWKPKDVYKNRSIELVQQQNVQLSLTEATEKAKQEFEKAGKDFLVETIKLGKLLRPNHLWGYYLFPDCYNHHYKKPGYNGSCFNVEIKRNDDLSWLWNESTALYPSIYLNTQQSPVAATLYVRNRVREAIRVSKIPDAKSPLPVFAYTRIVFTDQVLKFLSQDELVYTFGETVALGASGIVIWGT WENT R TKE SC QAIKE YM D T T LNPYIINVTLAAKMCSQVLCQEQGVCIRKNWNSSDYLHLNPDNFAIQLEKGGKFTVRGKPTLEDLEQFSEKFYCSCYSTLSCKEKADVKDTDAVDVCIADGVCIDAFLKPP HM36 34 Fourteen amino acid residues were substituted as L342W, S343E, I344N, M345T, S347T, M348K, K349E, L352Q, L353A, L354I, D355K, N356E, E359D and I361T, and before the amino-terminal residue F38 and at The carboxy-terminal residue M473 was then cleaved. FRAPPVIPNVPFLWAWNAPSEFCLGKFDEPLDMSLFSFIGSPRINATGQGVTIFYVDRLGYYPYIDSITGVTVNGGIPQKISLQDHLDKAKKDITFYMPVDNLGMAVIDWEEWRPTWARNWKPKDVYKNRSIELVQQQNVQLSLTEATEKAKQEFEKAGKDFLVETIKLGKLLRPNHLWGYYLFPDCYNHHYKKPGYNGSCFNVEIKRNDDLSWLWNESTALYPSIYLNTQQSPVAATLYVRNRVREAIRVSKIPDAKSPLPVFAYTRIVFTDQVLKFLSQDELVYTFGETVALGASGIVIWGT WENT R TKE SC QAIKE YM D T T LNPYIINVTLAAKMCSQVLCQEQGVCIRKNWNSSDYLHLNPDNFAIQLEKGGKFTVRGKPTLEDLEQFSEKFYCSCYSTLSCKEKADVKDTDAVDVCIADGVCIDAFLKPPM HM37 35 Fourteen amino acid residues were substituted as L342W, S343E, I344N, M345T, S347T, M348K, K349E, L352Q, L353A, L354I, D355K, N356E, E359D and I361T, and before the amino-terminal residue F38 and at The carboxy-terminal residue E474 was followed by cleavage. FRAPPVIPNVPFLWAWNAPSEFCLGKFDEPLDMSLFSFIGSPRINATGQGVTIFYVDRLGYYPYIDSITGVTVNGGIPQKISLQDHLDKAKKDITFYMPVDNLGMAVIDWEEWRPTWARNWKPKDVYKNRSIELVQQQNVQLSLTEATEKAKQEFEKAGKDFLVETIKLGKLLRPNHLWGYYLFPDCYNHHYKKPGYNGSCFNVEIKRNDDLSWLWNESTALYPSIYLNTQQSPVAATLYVRNRVREAIRVSKIPDAKSPLPVFAYTRIVFTDQVLKFLSQDELVYTFGETVALGASGIVIWGT WENT R TKE SC QAIKE YM D T T LNPYIINVTLAAKMCSQVLCQEQGVCIRKNWNSSDYLHLNPDNFAIQLEKGGKFTVRGKPTLEDLEQFSEKFYCSCYSTLSCKEKADVKDTDAVDVCIADGVCIDAFLKPPME HM38 36 Fourteen amino acid residues were substituted as L342W, S343E, I344N, M345T, S347T, M348K, K349E, L352Q, L353A, L354I, D355K, N356E, E359D and I361T, and before the amino-terminal residue F38 and at The carboxy-terminal residue T475 was then cleaved. FRAPPVIPNVPFLWAWNAPSEFCLGKFDEPLDMSLFSFIGSPRINATGQGVTIFYVDRLGYYPYIDSITGVTVNGGIPQKISLQDHLDKAKKDITFYMPVDNLGMAVIDWEEWRPTWARNWKPKDVYKNRSIELVQQQNVQLSLTEATEKAKQEFEKAGKDFLVETIKLGKLLRPNHLWGYYLFPDCYNHHYKKPGYNGSCFNVEIKRNDDLSWLWNESTALYPSIYLNTQQSPVAATLYVRNRVREAIRVSKIPDAKSPLPVFAYTRIVFTDQVLKFLSQDELVYTFGETVALGASGIVIWGT WENT R TKE SC QAIKE YM D T T LNPYIINVTLAAKMCSQVLCQEQGVCIRKNWNSSDYLHLNPDNFAIQLEKGGKFTVRGKPTLEDLEQFSEKFYCSCYSTLSCKEKADVKDTDAVDVCIADGVCIDAFLKPPMET HM39 37 Fourteen amino acid residues were substituted as L342W, S343E, I344N, M345T, S347T, M348K, K349E, L352Q, L353A, L354I, D355K, N356E, E359D and I361T, and before the amino-terminal residue F38 and at The carboxy-terminal residue E476 was followed by cleavage. FRAPPVIPNVPFLWAWNAPSEFCLGKFDEPLDMSLFSFIGSPRINATGQGVTIFYVDRLGYYPYIDSITGVTVNGGIPQKISLQDHLDKAKKDITFYMPVDNLGMAVIDWEEWRPTWARNWKPKDVYKNRSIELVQQQNVQLSLTEATEKAKQEFEKAGKDFLVETIKLGKLLRPNHLWGYYLFPDCYNHHYKKPGYNGSCFNVEIKRNDDLSWLWNESTALYPSIYLNTQQSPVAATLYVRNRVREAIRVSKIPDAKSPLPVFAYTRIVFTDQVLKFLSQDELVYTFGETVALGASGIVIWGT WENT R TKE SC QAIKE YM D T T LNPYIINVTLAAKMCSQVLCQEQGVCIRKNWNSSDYLHLNPDNFAIQLEKGGKFTVRGKPTLEDLEQFSEKFYCSCYSTLSCKEKADVKDTDAVDVCIADGVCIDAFLKPPMETE HM40 38 Eleven amino acid residues were substituted as M345T, S347T, M348K, K349E, L352Q, L353A, L354I, D355K, N356E, E359D, and I361T, and were cleaved before the amino-terminal residue N37. NFRAPPVIPNVPFLWAWNAPSEFCLGKFDEPLDMSLFSFIGSPRINATGQGVTIFYVDRLGYYPYIDSITGVTVNGGIPQKISLQDHLDKAKKDITFYMPVDNLGMAVIDWEEWRPTWARNWKPKDVYKNRSIELVQQQNVQLSLTEATEKAKQEFEKAGKDFLVETIKLGKLLRPNHLWGYYLFPDCYNHHYKKPGYNGSCFNVEIKRNDDLSWLWNESTALYPSIYLNTQQSPVAATLYVRNRVREAIRVSKIPDAKSPLPVFAYTRIVFTDQVLKFLSQDELVYTFGETVALGASGIVIWGTLSI T R TKE SC QAIKE YM D T T LNPYIINVTLAAKMCSQVLCQEQGVCIRKNWNSSDYLHLNPDNFAIQLEKGGKFTVRGKPTLEDLEQFSEKFYCSCYSTLSCKEKADVKDTDAVDVCIADGVCIDAFLKPPMETEEPQIFYNASPSTLS HM41 39 Eleven amino acid residues were substituted as M345T, S347T, M348K, K349E, L352Q, L353A, L354I, D355K, N356E, E359D, and I361T, and were cleaved before the amino-terminal residue R39. RAPPVIPNVPFLWAWNAPSEFCLGKFDEPLDMSLFSFIGSPRINATGQGVTIFYVDRLGYYPYIDSITGVTVNGGIPQKISLQDHLDKAKKDITFYMPVDNLGMAVIDWEEWRPTWARNWKPKDVYKNRSIELVQQQNVQLSLTEATEKAKQEFEKAGKDFLVETIKLGKLLRPNHLWGYYLFPDCYNHHYKKPGYNGSCFNVEIKRNDDLSWLWNESTALYPSIYLNTQQSPVAATLYVRNRVREAIRVSKIPDAKSPLPVFAYTRIVFTDQVLKFLSQDELVYTFGETVALGASGIVIWGTLSI T R TKE SC QAIKE YM D T T LNPYIINVTLAAKMCSQVLCQEQGVCIRKNWNSSDYLHLNPDNFAIQLEKGGKFTVRGKPTLEDLEQFSEKFYCSCYSTLSCKEKADVKDTDAVDVCIADGVCIDAFLKPPMETEEPQIFYNASPSTLS HM42 40 Eleven amino acid residues were substituted as M345T, S347T, M348K, K349E, L352Q, L353A, L354I, D355K, N356E, E359D, and I361T, and were cleaved before the amino-terminal residue P41. PPVIPNVPFLWAWNAPSEFCLGKFDEPLDMSLFSFIGSPRINATGQGVTIFYVDRLGYYPYIDSITGVTVNGGIPQKISLQDHLDKAKKDITFYMPVDNLGMAVIDWEEWRPTWARNWKPKDVYKNRSIELVQQQNVQLSLTEATEKAKQEFEKAGKDFLVETIKLGKLLRPNHLWGYYLFPDCYNHHYKKPGYNGSCFNVEIKRNDDLSWLWNESTALYPSIYLNTQQSPVAATLYVRNRVREAIRVSKIPDAKSPLPVFAYTRIVFTDQVLKFLSQDELVYTFGETVALGASGIVIWGTLSI T R TKE SC QAIKE YM D T T LNPYIINVTLAAKMCSQVLCQEQGVCIRKNWNSSDYLHLNPDNFAIQLEKGGKFTVRGKPTLEDLEQFSEKFYCSCYSTLSCKEKADVKDTDAVDVCIADGVCIDAFLKPPMETEEPQIFYNASPSTLS HM43 41 Fourteen amino acid residues were substituted as L342W, S343E, I344N, M345T, S347T, M348K, K349E, L352Q, L353A, L354I, D355K, N356E, E359D and I361T, and before the amino-terminal residue F38 and at The carboxy-terminal residue I465 was followed by cleavage. FRAPPVIPNVPFLWAWNAPSEFCLGKFDEPLDMSLFSFIGSPRINATGQGVTIFYVDRLGYYPYIDSITGVTVNGGIPQKISLQDHLDKAKKDITFYMPVDNLGMAVIDWEEWRPTWARNWKPKDVYKNRSIELVQQQNVQLSLTEATEKAKQEFEKAGKDFLVETIKLGKLLRPNHLWGYYLFPDCYNHHYKKPGYNGSCFNVEIKRNDDLSWLWNESTALYPSIYLNTQQSPVAATLYVRNRVREAIRVSKIPDAKSPLPVFAYTRIVFTDQVLKFLSQDELVYTFGETVALGASGIVIWGS WENT R TKE SC QAIKE YM D T T LNPYIINVTLAAKMCSQVLCQEQGVCIRKNWNSSDYLHLNPDNFAIQLEKGGKFTVRGKPTLEDLEQFSEKFYCSCYSTLSCKEKADVKDTDAVDVCIADGVCI HM44 42 Fourteen amino acid residues were substituted as L342W, S343E, I344N, M345T, S347T, M348K, K349E, L352Q, L353A, L354I, D355K, N356E, E359D and I361T, and before the amino-terminal residue F38 and and Cleavage occurs after the carboxy-terminal residue D466. FRAPPVIPNVPFLWAWNAPSEFCLGKFDEPLDMSLFSFIGSPRINATGQGVTIFYVDRLGYYPYIDSITGVTVNGGIPQKISLQDHLDKAKKDITFYMPVDNLGMAVIDWEEWRPTWARNWKPKDVYKNRSIELVQQQNVQLSLTEATEKAKQEFEKAGKDFLVETIKLGKLLRPNHLWGYYLFPDCYNHHYKKPGYNGSCFNVEIKRNDDLSWLWNESTALYPSIYLNTQQSPVAATLYVRNRVREAIRVSKIPDAKSPLPVFAYTRIVFTDQVLKFLSQDELVYTFGETVALGASGIVIWGS WENT R TKE SC QAIKE YM D T T LNPYIINVTLAAKMCSQVLCQEQGVCIRKNWNSSDYLHLNPDNFAIQLEKGGKFTVRGKPTLEDLEQFSEKFYCSCYSTLSCKEKADVKDTDAVDVCIADGVCID HM45 43 Fourteen amino acid residues were substituted as L342W, S343E, I344N, M345T, S347T, M348K, K349E, L352Q, L353A, L354I, D355K, N356E, E359D and I361T, and before the amino-terminal residue F38 and and Cleavage occurs after the carboxy-terminal residue A467. FRAPPVIPNVPFLWAWNAPSEFCLGKFDEPLDMSLFSFIGSPRINATGQGVTIFYVDRLGYYPYIDSITGVTVNGGIPQKISLQDHLDKAKKDITFYMPVDNLGMAVIDWEEWRPTWARNWKPKDVYKNRSIELVQQQNVQLSLTEATEKAKQEFEKAGKDFLVETIKLGKLLRPNHLWGYYLFPDCYNHHYKKPGYNGSCFNVEIKRNDDLSWLWNESTALYPSIYLNTQQSPVAATLYVRNRVREAIRVSKIPDAKSPLPVFAYTRIVFTDQVLKFLSQDELVYTFGETVALGASGIVIWGS WENT R TKE SC QAIKE YM D T T LNPYIINVTLAAKMCSQVLCQEQGVCIRKNWNSSDYLHLNPDNFAIQLEKGGKFTVRGKPTLEDLEQFSEKFYCSCYSTLSCKEKADVKDTDAVDVCIADGVCIDA HP46 44 Fifteen amino acid residues were substituted as T341S, L342W, S343E, I344N, M345T, S347T, M348K, K349E, L352Q, L353A, L354I, D355K, N356E, E359D, and I361T, and before the amino-terminal residue F38 and cleavage after the carboxy-terminal residue F468. FRAPPVIPNVPFLWAWNAPSEFCLGKFDEPLDMSLFSFIGSPRINATGQGVTIFYVDRLGYYPYIDSITGVTVNGGIPQKISLQDHLDKAKKDITFYMPVDNLGMAVIDWEEWRPTWARNWKPKDVYKNRSIELVQQQNVQLSLTEATEKAKQEFEKAGKDFLVETIKLGKLLRPNHLWGYYLFPDCYNHHYKKPGYNGSCFNVEIKRNDDLSWLWNESTALYPSIYLNTQQSPVAATLYVRNRVREAIRVSKIPDAKSPLPVFAYTRIVFTDQVLKFLSQDELVYTFGETVALGASGIVIWG SWENT R TKE SC QAIKE YM D T T LNPYIINVTLAAKMCSQVLCQEQGVCIRKNWNSSDYLHLNPDNFAIQLEKGGKFTVRGKPTLEDLEQFSEKFYCSCYSTLSCKEKADVKDTDAVDVCIADGVCIDAF HM47 45 Fourteen amino acid residues were substituted as L342W, S343E, I344N, M345T, S347T, M348K, K349E, L352Q, L353A, L354I, D355K, N356E, E359D and I361T, and before the amino-terminal residue F38 and at The carboxy-terminal residue P478 was then cleaved. FRAPPVIPNVPFLWAWNAPSEFCLGKFDEPLDMSLFSFIGSPRINATGQGVTIFYVDRLGYYPYIDSITGVTVNGGIPQKISLQDHLDKAKKDITFYMPVDNLGMAVIDWEEWRPTWARNWKPKDVYKNRSIELVQQQNVQLSLTEATEKAKQEFEKAGKDFLVETIKLGKLLRPNHLWGYYLFPDCYNHHYKKPGYNGSCFNVEIKRNDDLSWLWNESTALYPSIYLNTQQSPVAATLYVRNRVREAIRVSKIPDAKSPLPVFAYTRIVFTDQVLKFLSQDELVYTFGETVALGASGIVIWGT WENT R TKE SC QAIKE YM D T T LNPYIINVTLAAKMCSQVLCQEQGVCIRKNWNSSDYLHLNPDNFAIQLEKGGKFTVRGKPTLEDLEQFSEKFYCSCYSTLSCKEKADVKDTDAVDVCIADGVCIDAFLKPPMETEEP HM48 46 Fourteen amino acid residues were substituted as L342W, S343E, I344N, M345T, S347T, M348K, K349E, L352Q, L353A, L354I, D355K, N356E, E359D and I361T, and before the amino-terminal residue F38 and at The carboxy-terminal residue I480 was followed by cleavage. FRAPPVIPNVPFLWAWNAPSEFCLGKFDEPLDMSLFSFIGSPRINATGQGVTIFYVDRLGYYPYIDSITGVTVNGGIPQKISLQDHLDKAKKDITFYMPVDNLGMAVIDWEEWRPTWARNWKPKDVYKNRSIELVQQQNVQLSLTEATEKAKQEFEKAGKDFLVETIKLGKLLRPNHLWGYYLFPDCYNHHYKKPGYNGSCFNVEIKRNDDLSWLWNESTALYPSIYLNTQQSPVAATLYVRNRVREAIRVSKIPDAKSPLPVFAYTRIVFTDQVLKFLSQDELVYTFGETVALGASGIVIWGT WENT R TKE SC QAIKE YM D T T LNPYIINVTLAAKMCSQVLCQEQGVCIRKNWNSSDYLHLNPDNFAIQLEKGGKFTVRGKPTLEDLEQFSEKFYCSCYSTLSCKEKADVKDTDAVDVCIADGVCIDAFLKPPMETEEPQI HM49 47 Fourteen amino acid residues were substituted as L342W, S343E, I344N, M345T, S347T, M348K, K349E, L352Q, L353A, L354I, D355K, N356E, E359D and I361T, and before the amino-terminal residue F38 and at The carboxy-terminal residue Y482 was then cleaved. FRAPPVIPNVPFLWAWNAPSEFCLGKFDEPLDMSLFSFIGSPRINATGQGVTIFYVDRLGYYPYIDSITGVTVNGGIPQKISLQDHLDKAKKDITFYMPVDNLGMAVIDWEEWRPTWARNWKPKDVYKNRSIELVQQQNVQLSLTEATEKAKQEFEKAGKDFLVETIKLGKLLRPNHLWGYYLFPDCYNHHYKKPGYNGSCFNVEIKRNDDLSWLWNESTALYPSIYLNTQQSPVAATLYVRNRVREAIRVSKIPDAKSPLPVFAYTRIVFTDQVLKFLSQDELVYTFGETVALGASGIVIWGT WENT R TKE SC QAIKE YM D T T LNPYIINVTLAAKMCSQVLCQEQGVCIRKNWNSSDYLHLNPDNFAIQLEKGGKFTVRGKPTLEDLEQFSEKFYCSCYSTLSCKEKADVKDTDAVDVCIADGVCIDAFLKPPMETEEPQIFY HM50 48 Fourteen amino acid residues were substituted as L342W, S343E, I344N, M345T, S347T, M348K, K349E, L352Q, L353A, L354I, D355K, N356E, E359D and I361T, and before the amino-terminal residue F38 and and Cleavage occurs after the carboxy-terminal residue A484. FRAPPVIPNVPFLWAWNAPSEFCLGKFDEPLDMSLFSFIGSPRINATGQGVTIFYVDRLGYYPYIDSITGVTVNGGIPQKISLQDHLDKAKKDITFYMPVDNLGMAVIDWEEWRPTWARNWKPKDVYKNRSIELVQQQNVQLSLTEATEKAKQEFEKAGKDFLVETIKLGKLLRPNHLWGYYLFPDCYNHHYKKPGYNGSCFNVEIKRNDDLSWLWNESTALYPSIYLNTQQSPVAATLYVRNRVREAIRVSKIPDAKSPLPVFAYTRIVFTDQVLKFLSQDELVYTFGETVALGASGIVIWGT WENT R TKE SC QAIKE YM D T T LNPYIINVTLAAKMCSQVLCQEQGVCIRKNWNSSDYLHLNPDNFAIQLEKGGKFTVRGKPTLEDLEQFSEKFYCSCYSTLSCKEKADVKDTDAVDVCIADGVCIDAFLKPPMETEEPQIFYNA HM51 49 Fourteen amino acid residues were substituted as L342W, S343E, I344N, M345T, S347T, M348K, K349E, L352Q, L353A, L354I, D355K, N356E, E359D and I361T, and before the amino-terminal residue F38 and at The carboxy-terminal residue P486 was then cleaved. FRAPPVIPNVPFLWAWNAPSEFCLGKFDEPLDMSLFSFIGSPRINATGQGVTIFYVDRLGYYPYIDSITGVTVNGGIPQKISLQDHLDKAKKDITFYMPVDNLGMAVIDWEEWRPTWARNWKPKDVYKNRSIELVQQQNVQLSLTEATEKAKQEFEKAGKDFLVETIKLGKLLRPNHLWGYYLFPDCYNHHYKKPGYNGSCFNVEIKRNDDLSWLWNESTALYPSIYLNTQQSPVAATLYVRNRVREAIRVSKIPDAKSPLPVFAYTRIVFTDQVLKFLSQDELVYTFGETVALGASGIVIWGT WENT R TKE SC QAIKE YM D T T LNPYIINVTLAAKMCSQVLCQEQGVCIRKNWNSSDYLHLNPDNFAIQLEKGGKFTVRGKPTLEDLEQFSEKFYCSCYSTLSCKEKADVKDTDAVDVCIADGVCIDAFLKPPMETEEPQIFYNASP HM52 50 Fourteen amino acid residues were substituted as L342W, S343E, I344N, M345T, S347T, M348K, K349E, L352Q, L353A, L354I, D355K, N356E, E359D and I361T, and before the amino-terminal residue F38 and at The carboxy-terminal residue T488 was then cleaved. FRAPPVIPNVPFLWAWNAPSEFCLGKFDEPLDMSLFSFIGSPRINATGQGVTIFYVDRLGYYPYIDSITGVTVNGGIPQKISLQDHLDKAKKDITFYMPVDNLGMAVIDWEEWRPTWARNWKPKDVYKNRSIELVQQQNVQLSLTEATEKAKQEFEKAGKDFLVETIKLGKLLRPNHLWGYYLFPDCYNHHYKKPGYNGSCFNVEIKRNDDLSWLWNESTALYPSIYLNTQQSPVAATLYVRNRVREAIRVSKIPDAKSPLPVFAYTRIVFTDQVLKFLSQDELVYTFGETVALGASGIVIWGT WENT R TKE SC QAIKE YM D T T LNPYIINVTLAAKMCSQVLCQEQGVCIRKNWNSSDYLHLNPDNFAIQLEKGGKFTVRGKPTLEDLEQFSEKFYCSCYSTLSCKEKADVKDTDAVDVCIADGVCIDAFLKPPMETEEPQIFYNASPST

Meanwhile, previous studies reported that the enzymatic activity of wild-type PH20 was altered depending on the excised position of the carboxy-terminal amino acid residue. However, in the present invention, the specific α-helix forming the secondary structure of PH20 is replaced by the α-helix of human hyaluronidase, thereby constructing a PH20 variant having higher stability than wild-type PH20, and In these variants, the interaction between the substituted α-helical domain and other secondary structures of PH20 appears to be in a pattern different from that of wild-type PH20, such that the variant has a certain A degree or higher enzyme activity.

Furthermore, in the present invention, an attempt was made to increase the expression of recombinant PH20 protein by replacing the original human PH20 message peptide with a message peptide of another protein that exhibits a high degree of protein expression in animal cells.

Therefore, in another embodiment, the PH20 variant contained in the pharmaceutical composition according to the present invention may comprise a human hyaluronidase-1 (human hyaluronidase-1, Hyal1), human growth hormone or human hyaluronidase at its amino terminal. The message peptide of serum albumin, replaces the message peptide of the wild-type PH20 that is made up of M1 to T35, and preferably can comprise the message peptide that is derived from human growth hormone with the amino acid sequence of MATGSRTSLLLAFGLLCLPWLQEGSA according to SEQ ID NO:2 , a message peptide derived from human serum albumin having the amino acid sequence of MKWVTFISLLFLFSSAYS according to SEQ ID NO: 3 or a message peptide derived from human Hyal1 having the amino acid sequence of MAAHLLPICALFLTLLDMAQG according to SEQ ID NO: 4, such as shown in Table 5, but the present invention is not limited thereto.

Table 5, the amino acid sequence of the message peptide of human growth hormone, human serum albumin or human Hyal1 Source of message peptides amino acid sequence SEQ ID NO: human growth hormone MATGSRTSLLLAFGLLCLPWLQEGSA 2 human serum albumin MKWVTFISLLFLFSSAYS 3 humanHyal1 MAAHLLPICALFLTLLDMAQG 4

Among the PH20 variants contained in the pharmaceutical composition according to the present invention, the variant with a histidine tag (6xHis-tag) attached to the carboxy terminus is named HM, and the variant without a histidine tag is named HP. In addition, the mature wild-type PH20 (L36-S490) with a histidine tag attached to the carboxy-terminus was named WT, and the mature wild-type PH20 (L36 to Y482 ) named HW2.

HP46 (SEQ ID NO: 44) is a PH20 variant obtained by modeling the protein structure (Chao et al., 2007), then replace the amino acid sequence of the amino acid sequence of α-helix 8 and the linking domain between α-helix 7 and α-helix 8 with the amino acid sequence of Hyal1, and make the amino terminal at F38 was cleaved and the carboxy terminus was cleaved after F468. In particular, α-helix 8 is located outside the protein tertiary structure of PH20 and has fewer interactions with adjacent α-helices or β-strands than other α-helices. Generally speaking, there is a trade-off relationship between enzyme activity and thermal stability, so the higher the thermal stability of the protein, the lower the enzyme activity, and when the enzyme activity is due to the flexibility of the protein structure ) improves and increases, the thermal stability tends to decrease. However, the specific activity of HP46 measured by Turbidimetric assay at pH 7 was about 46 units/μg, which was estimated to be about twice that of wild-type PH20 (about 23 units/μg).

The thermal stability of proteins can be evaluated based on the melting temperature (Tm) at which 50% of the protein's tertiary structure is denatured (denature) and the aggregation temperature (Tagg) at which aggregation occurs between proteins. In general, the aggregation temperature of proteins tends to be lower than their melting point. Compared with the α-helix 8 of PH20, the α-helix 8 of Hyal1 exhibits higher hydrophilicity. The α-helix 8 of the substituted Hyal1 increases the hydrophilicity of the protein surface of HP46, thereby delaying the aggregation effect between proteins due to the hydrophobic effect, so the aggregation temperature is 51°C, which is compared with the aggregation of wild-type PH20 For temperature (46.5°C), an increase of 4.5°C was observed.

HP46 is a variant with substituted amino acid residues in α-helix 8 and the linking domain between α-helix 7 and α-helix 8, wherein T341 is substituted with serine. When amino acid residue 341 was threonine, the enzyme activity was similar to that of wild-type PH20, but when it was replaced by serine, the enzyme activity increased about 2-fold, and it was confirmed that even in the substrate gel analysis (substrate gel assay), the resulting variant hydrolyzed 5 to 6 times more hyaluronic acid compared to wild-type PH20. Substrate gel analysis involves the process of protein denaturation and refolding, which means that the refolding and restoration of the protein tertiary structure of HP46 is enhanced compared with wild-type PH20.

The amount of PH20 variant in the pharmaceutical composition according to the present invention is at least 50 units/ml, preferably in the range of 100 units/ml to 20,000 units/ml, more preferably about 150 units/ml to about In the range of 18,000 units/ml, more preferably in the range of 1,000 units/ml to 16,000 units/ml, most preferably in the range of 1,500 units/ml to 12,000 units/ml.

Examples of drugs included in pharmaceutical compositions according to the present invention include, but are not limited to, protein drugs, antibody drugs, small molecules, aptamers, RNAi, antisense RNA, and cellular therapeutics, such as chimeric antigen receptor T cells ( chimeric antigen receptor T-cell, CAR-T) or chimeric antigen receptor natural killer cells (CAR-natural killer, CAR-NK), which can not only use currently available drugs on the market, but also can be used in clinical trials or drugs in development.

As drugs, protein drugs or antibody drugs are preferably used.

The "protein drug" contained in the pharmaceutical composition of the present invention is a drug composed of amino acids, so it exhibits the effect of treating or preventing diseases through the activity of the protein, and is a drug composed of proteins different from antibody drugs , and can be selected from cytokines, therapeutic enzymes, hormones, soluble receptors and their fusion proteins, insulin or its analogs, bone morphogenic protein (bone morphogenetic protein, BMP), erythropoietin (erythropoietin) and serum-derived protein (serum-derived protein), but the present invention is not limited thereto.

The cytokines contained in the pharmaceutical composition according to the present invention can be selected from interferon (interferon), interleukin (interleukin), colony-stimulating factor (colony-stimulating factor, CSF), tumor necrosis factor (tumor necrosis factor, TNF). ) and tissue growth factor (tissue growth factor, TGF), but the present invention is not limited thereto.

Therapeutic enzymes may include, but are not limited to, β-glucocerebrosidase and agalsidase β.

The fusion protein of the extracellular domain (extracellular domain) of the receptor of the soluble receptor contained in the pharmaceutical composition of the present invention is the Fc region of an antibody or the like fused to the protein of the soluble receptor. Soluble receptors are soluble forms of receptors to which ligands associated with diseases bind, examples of which include forms in which the Fc region is fused to TNF-α soluble receptors (such as products comprising etanercept components and forms similar thereto), Forms in which the Fc region is fused to vascular endothelial cell growth factor (VEGF) soluble receptors (e.g., products comprising alefacept components and forms similar thereto), forms in which the Fc region is fused to CTLA-4 (e.g., forms comprising abatacept or belatacept components and similar forms) products in its form), forms in which the Fc region is fused to the interleukin-1 soluble receptor (such as products containing rilonacept components and products similar to it), and forms in which the Fc region is fused to LFA3 soluble receptors (such as containing alefacept components and similar to its form), but the invention is not limited thereto.

The hormone contained in the pharmaceutical composition according to the present invention refers to the hormone or its analogue injected into the human body for the treatment or prevention of diseases caused by hormone deficiency, etc. Examples include, but are not limited to, human growth hormone, Estrogen and progesterone.

The serum-derived protein contained in the pharmaceutical composition according to the present invention is a protein present in blood plasma, and includes proteins extracted from blood plasma and recombinant proteins produced, examples of which may include, but are not limited to, fibrinogen (fibrinogen), von Willebrand factor, albumin, thrombin, factor II (FII), factor V (FV), factor VII (FVII), factor IX (FIX), factor X (FX), and factor (FXI) .

The antibody drug included in the pharmaceutical composition of the present invention can be a monoclonal antibody drug or a polyclonal antibody drug.

The monoclonal antibody drug according to the present invention is a protein comprising monoclonal antibodies and monoclonal antibody fragments, wherein the monoclonal antibodies and monoclonal antibody fragments can specifically bind to antigens associated with specific diseases. Monoclonal antibodies also include bispecific antibodies, and proteins including monoclonal antibodies or fragments thereof conceptually include antibody-drug conjugates (ADCs).

Examples of antigens associated with specific diseases include 4-1BB, integrin, amyloid beta, angiopoietin (angiopoietin 1 or 2), angiopoietin analog 3, B cell activating factor (B-cell-activating factor, BAFF), B7-H3, complement 5 (complement 5), CCR4, CD3, CD4, CD6, CD11a, CD19, CD20, CD22, CD30, CD33, CD38, CD52, CD62, CD79b, CD80, CGRP, claudin-18 (Claudin-18), complement factor D (complement factor D), CTLA4, DLL3, EGF receptor, hemophilia factor (hemophilia factor), Fc receptor, FGF23 , folate receptor, GD2, GM-CSF, HER2, HER3, interferon receptor, interferon gamma, IgE, IGF-1 receptor, interleukin 1, interleukin 2 receptor, interleukin IL-4 receptor, IL-5, IL-5 receptor, IL-6, IL-6 receptor, IL-7, IL-12/23, IL-13, IL-17A , Interleukin 17 receptor A, Interleukin 31 receptor, Interleukin 36 receptor, LAG3, LFA3, NGF, PVSK9, PD-1, PD-L1, RANK-L, SLAMF7, tissue factor, TNF, VEGF, VEGF receptor and von Willebrand factor (vWF), but the present invention is not limited thereto.

The following are, but are not limited to, proteins comprising monoclonal antibodies or monoclonal antibody fragments associated with specific diseases: utomilumab as an anti-4-1BB antibody; natalizumab, etrolizumab, vedolizumab, and bimagrumab as antibodies against integrins; bapineuzumab, crenezumab, solanezumab, aducanumab, and gantenerumab as antibodies against amyloid beta; Antibodies against angiogenin, such as AMG780, MEDI 3617 against angiogenin 1 and 2, nesvacumab against angiogenin 2, and vanucizumab which is a bispecific antibody against angiogenin 2 and VEGF; Evinacumab as an antibody against angiogenin analogue 3; tabalumab, lanalumab, and belimumab as antibodies against cell activating factor (BAFF); omburtamab as an antibody to B7-H3; Ravulizumab and eculizumab as antibodies against complement 5; mogamulizumab as an antibody against CCR4; otelixizumab, teplizumab and muromonab are antibodies against CD3, tebentafusp is a bispecific antibody against GP100 and CD3, blinatumomab is a bispecific antibody against CD19 and CD3, and REGN1979 is a bispecific antibody against CD20 and CD3; ibalizumab and zanolimumab as antibodies against CD4; itolizumab as an antibody against CD6; efalizumab as an antibody against CD11a; inebilizumab, tafasitamab and loncastuximab tesirine (which is an antibody-drug complex) as antibodies against CD19; ocrelizumab, ublituximab, obinutuzumab, ofatumumab, rituximab, tositumomab, and ibritumomab tiuxetan (which is an antibody-drug complex) as antibodies against CD20; epratuzumab, inotuzumab ozogamicin (which is an antibody-drug complex) and moxetumomab pasudotox as antibodies against CD22; brentuximab vedotin as an antibody-drug complex against CD30; vadastuximab talirine and gemtuzumab ozogamicin as an antibody-drug complex against CD33; daratumumab and isatuximab as antibody-drug complexes targeting CD38; alemtuzumab as an antibody against CD52; crizanlizumab as an antibody against CD62; polaruzumab vedotin as an antibody-drug complex against CD79b; galiximab as an antibody against CD80; eptinezumab, fremanezumab, galcanezumab and erenumab as antibodies against CGRP; zolbetuximab as an antibody against claudin-18; lampalizumab as an antibody against complement factor D; tremelimumab, zalifrelimab and ipilimumab as antibodies against CTLA4; rovalpituzumab tesirine as an antibody-drug complex targeting DLL3; cetuximab, depatuxizumab, zalutumumab, necitumumab, and panitumumab as antibodies against the EGF receptor; emicizumab as a bispecific antibody against coagulation factor IX and factor X (which is hemophilia factor); nipocalimab and rozanolixizumab as antibodies against Fc receptors; Burosumab as an antibody against FGF23; farletuzumab as an antibody against the folate receptor, and mirvetuximab soravtansine as an antibody-drug complex against the folate receptor; dinutuximab and naxitamab as antibodies against GD2; otilimab as an antibody against GM-CSF; margetuximab, pertuzumab, and trastuzumab as antibodies against HER2, and trastuzumab deruxtecan, trastuzumab emtansine, and trastuzumab duocarmazine as antibody-drug complexes against HER2; patritumab as an antibody against HER3; anifrolumab as an antibody against interferon receptors; emapalumab as an antibody against interferon gamma; Ligelizumab and omalizumab as antibodies against IgE; Dalotuzumab, figitumumab and teprotumumab as antibodies against IGF-1 receptor; gebokizumab and canakinumab as antibodies against interleukin 1; Daclizumab and basiliximab as antibodies against interleukin-2 receptor; dupilumab as an antibody against the interleukin-4 receptor; Mepolizumab and reslizumab as antibodies against interleukin 5; benralizumab as an antibody against the interleukin-5 receptor; clazakizumab, olokizumab, sirukumab, and siltuximab as antibodies against interleukin-6; Sarilumab, satralizumab, tocilizumab and REGN88 as antibodies against interleukin-6 receptor; secukinumab as an antibody against interleukin 7; Ustekinumab and briakinumab as antibodies against interleukin 12/23; lebrikizumab and tralokinumab as antibodies against interleukin-13; Ixekizumab and bimekizumab as antibodies against interleukin 17A; Brodalumab as an antibody against interleukin 17 receptor A; brazikumab, guselkumab, risankizumab, tildrakizumab, and mirikizumab as antibodies against interleukin-23; nemolizumab as an antibody against the interleukin-31 receptor; spesolimab as an antibody against the interleukin-36 receptor; relatlimab as an antibody against LAG3; narsoplimab as an antibody against NASP2; Fasinumab and tanezumab as antibodies against NGF; alirocumab, evolocumab and bococizumab as antibodies against PVSK9; Lambrolizumab, balstilimab, camrelizumab, cemiplimab, dostarlimab, prolgolimab, shintilimab, spartalizumab, tislelizumab, pembrolizumab and nivolumab as antibodies against PD-1; atezolizumab, avelumab, envafolimab and durvalumab as antibodies against PD-L1, and bintrafusp alpha as bispecific antibodies against TGF beta and PD-L1; denosumab as an antibody against RANK-L; elotuzumab as an antibody against SLAMF7; concizumab and marstacimab as antibodies against tissue factor; Antibodies against TNF, especially TNFα, including infliximab, adalimumab, golimumab, antibody fragments certolizumab pegol and ozoralizumab (which are bispecific antibodies against TNF and albumin); Antibodies against VEGF, including brolucizumab, ranibizumab, bevacizumab and faricimab (which are bispecific antibodies against VEGF and Ang2); ramucirumab as an antibody against the VEGF receptor; and Caplacizumab as an antibody against vWF

At the same time, overexpression of human epidermal growth factor receptor 2 (HER2) is observed in about 20-25% of breast cancer patients, which promotes cell division. Breast cancer with overexpression of HER2 progresses rapidly and is aggressive , and compared with breast cancer with low expression of HER2, the response to chemotherapy is low, so its prognosis (prognosis) is unfavorable. Trastuzumab is a monoclonal antibody drug targeting HER2, which specifically binds to HER2 on the surface of cancer cells overexpressing HER2 to inhibit the transmission of cell replication and proliferation, thereby slowing down the progress of tumors. Trastuzumab has been approved by the United States Food and Drug Administration (FDA) for the treatment of breast cancer in the United States in 1998 and by the Korean Food and Drug Administration (KFDA) in 2003. Since then, the efficacy of trastuzumab has also been recognized in HER2-overexpressing gastric cancer, and thus has been used as a therapeutic agent for gastric cancer.

Roche's Heaiping subcutaneous injection preparation (trade name: Herceptin) consists of 440 mg of trastuzumab as the main ingredient, and the freeze-dried trastuzumab is mixed with normal saline and injected into the vein. On the other hand, the subcutaneous injection preparation of trastuzumab (trade name: Heaiping SC) is a liquid preparation of 5 ml, and contains 600 mg (120 mg/ml) of trastuzumab as a main ingredient, and contains 20 mM histidine (pH 5.5), 210 mM trehalose, 10 mM methionine, 0.04% polysorbate 20 and 10,000 units of rHuPH20 (2,000 units/ml, 0.004%, 40 μg/ml) as additives.

The shelf life of Heaiping subcutaneous injection preparation is 21 months. The intravenous formulation of trastuzumab is in lyophilized form and has a shelf life of 30 months, but the subcutaneous formulation of trastuzumab is in liquid form and has a shorter shelf life of 21 months. For this reason, it can be estimated that the stability of one or more trastuzumab and recombinant human hyaluronidase PH20 in liquid formulations is limited.

Herein, in the present invention, from the point of view of the properties of the PH20 variant according to the present invention, compared to wild-type human hyaluronidase PH20 and recombinant human PH20 available from Halozyme, the PH20 variant not only has The increased enzyme activity also has a high protein aggregation temperature, so it exhibits enhanced thermal stability, and the shelf life of the subcutaneous injection formulation is determined to be long-term, preferably 21 months or longer.

In the pharmaceutical composition according to the present invention, the content of the antibody drug can be in the range of 5 mg/ml to 500 mg/ml, preferably 20 mg/ml to 200 mg/ml, more preferably 100 mg/ml to 150 mg/ml mg/ml, optimally 120 ± 18 mg/ml, for example about 110 mg/ml, about 120 mg/ml, or about 130 mg/ml.

The polyclonal antibody contained in the pharmaceutical composition according to the present invention is preferably serum antibody extracted from serum, such as immunoglobulin, but not limited thereto.

In the case of small molecule compounds, any drug requiring a rapid effect for treatment or prophylaxis can be used without limitation. For example, morphine-based painkillers can be used (Thomas et al., 2009). Furthermore, when used as therapeutic agents for tissue necrosis caused by anticancer drugs, small molecule compounds can be used alone or in combination with antidote drugs such as Vinca alkaloids and Taxanes Use (Kreidieh et al., 2016).

The pharmaceutical composition according to the present invention may further comprise one or more of the group consisting of buffer, stabilizer and surfactant.

The buffer contained in the pharmaceutical composition of the present invention can be used without limitation, as long as it can achieve pH 4 to 8, preferably 5 to 7, the buffer is preferably selected from malate, formate, citric acid Salt, acetate, propionate, pyridine, piperidine, cacodylate, succinate, 2-(N-metholino)ethanesulfonic acid (MES), histidine, trimethylolamino Methane (Tris), bis-(2-hydroxyethyl)aminotrishydroxymethylaminomethane (bis-Tris), phosphate, ethanolamine, carbonate, piperazine-N,N'-bis(2-ethyl Sulfonic acid) (PIPES), imidazole, 1,3-bis(tris(hydroxymethyl)methylamino)propane (BIS-TRIS propane), N,N-bis(2-hydroxyethyl)-2-amine One or more of the group consisting of ethyl ethanesulfonic acid (BES), 3-(N-𠰌linyl)propanesulfonic acid (MOPS), hydroxyethylpiper𠯤ethanesulfonic acid (HEPES), pyrophosphate and triethanolamine Or, preferably a histidine buffer, such as L-histidine/HCl, but not limited thereto.

The concentration of the buffer may range from 0.001 mM to 200 mM, preferably from 1 mM to 50 mM, more preferably from 5 mM to 40 mM, most preferably from 10 mM to 30 mM.

In the pharmaceutical composition according to the present invention, stabilizers can be used without limitation, as long as they are generally used in this field to stabilize proteins. Preferably, stabilizers can be selected from carbohydrates, sugars or hydrates thereof, One or more of the group consisting of sugar alcohol or its hydrate and amino acid.

Carbohydrates, sugars or sugar alcohols used as stabilizers can be selected from trehalose or its hydrate, sucrose, saccharin, glycerin, butylene glycol, threitol, xylitol, arabitol, ribitol, mannitol , sorbitol, galactitol, trehalitol, iditol, inositol, heptanol, isomalt, maltitol, polysaccharide alcohol, cyclodextrin, hydroxypropyl cyclodextrin and glucose One or more of the groups formed, but not limited to this.

The amino acid may be selected from the group consisting of glutamic acid, glutamic acid, glycine, lysine, lysilysine, leucine, methionine, valine, serine , Selenium Methionine, Citrulline, Arginine, Asparagine, Aspartic Acid, Ornithine, Isoleucine, Taurine, Theanine, Threonine, Tryptophan , one or more of the group consisting of tyrosine, phenylalanine, proline, pyrrolysine, histidine and alanine, but not limited thereto.

In the pharmaceutical composition according to the present invention, the concentration of sugar or sugar alcohol used as a stabilizer can be in the range of 0.001 mM to 500 mM, preferably 100 mM to 300 mM, more preferably 150 mM to 250 mM, most preferably Preferably 180 mM to 230 mM, specifically, may be about 210 mM.

In addition, in the pharmaceutical composition according to the present invention, the amino acid used as a stabilizer may range from 1 mM to 100 mM, preferably from 3 mM to 30 mM, more preferably from 5 mM to 25 mM, and most preferably It may range from 7 mM to 20 mM, specifically, from 8 mM to 15 mM.

The pharmaceutical composition according to the present invention may further comprise a surfactant.

Preferably, the surfactant can be a nonionic surfactant, such as polyoxyethylene-sorbitan fatty acid ester (polysorbate or Tween), polyethylene-polypropylene glycol, polyoxyethylene-stearate, poly Oxyethylene alkyl ethers (such as polyoxyethylene monolauryl ether, alkylphenyl polyoxyethylene ether [Triton-X] and polyoxyethylene-polyoxypropylene copolymer [Poloxamer and Pluronic]) and dodecyl sulfonate sodium phosphate (SDS), but not limited thereto.

More preferably, polysorbate can be used. The polysorbate may be polysorbate 20 or polysorbate 80, but is not limited thereto.

In the pharmaceutical composition according to the present invention, the concentration of the nonionic surfactant can be in the range of 0.0000001% (w/v) (weight/volume) to 0.5% (w/v), preferably 0.000001% (w/ v) to 0.4% (w/v), more preferably 0.00001% (w/v) to 0.3% (w/v), most preferably 0.001% (w/v) to 0.2% (w/v).

In one embodiment, the pharmaceutical composition according to the present invention may comprise 50-350 mg/ml of antibody (such as anti-HER2 antibody or immune checkpoint antibody), a histidine buffer solution with pH 5.5±2.0, 10-400 mM α,α-Trehalose, 1-50 mM methionine and 0.0000001% (w/v) to 0.5% (w/v) polysorbate.

In a more specific embodiment, the pharmaceutical composition according to the present invention may comprise 120 mg/ml of anti-HER2 antibody or immune checkpoint antibody, 20 mM amino acid buffer at pH 5.5±2.0, 210 mM α,α-marine Sugar, 10 mM methionine and 2,000 units/ml of PH20 variant, and may further contain 0.005% (w/v) to 0.1% (w/v) polysorbate.

The pharmaceutical composition according to the present invention can be administered by intravenous injection, subcutaneous injection, intramuscular injection, intraperitoneal injection, endothelial administration, topical administration, intrapulmonary administration, intrarectal administration and the like, subcutaneous The administration is preferably performed by subcutaneous injection, more preferably the pharmaceutical composition is used as an injectable preparation for subcutaneous injection.

Therefore, another embodiment of the present invention provides a preparation comprising the pharmaceutical composition according to the present invention, preferably an injection preparation for subcutaneous injection.

Injection formulations for subcutaneous injection are presented in a ready-to-inject form that requires no additional dilution process and can be presented in pre-filled syringes, glass ampoules or plastic containers.

The present invention also relates to a method of treating a disease using the pharmaceutical composition or preparation according to the present invention.

The diseases that can be treated with the pharmaceutical composition or preparation according to the present invention are not particularly limited, as long as they are diseases that can be treated with the drug combined with the PH20 variant according to the present invention.

Diseases that can be treated using the pharmaceutical composition or preparation according to the present invention may be cancer or autoimmune diseases, but are not limited thereto.

The cancer or malignant tumor (carcinoma) that can be treated with the pharmaceutical composition or preparation according to the present invention is not particularly limited, and includes both solid cancer and blood cancer. Examples of these cancers include skin cancer (such as melanoma), liver cancer, hepatocellular carcinoma, stomach cancer, breast cancer, lung cancer, ovarian cancer, bronchial cancer, nasopharyngeal cancer, laryngeal cancer, pancreatic cancer, bladder cancer, colon cancer Colorectal cancer, colon cancer, cervical cancer, brain cancer, prostate cancer, bone cancer, thyroid cancer, parathyroid cancer, kidney cancer, esophagus cancer, biliary tract cancer, testicular cancer, rectal cancer ( rectal cancer), head and neck cancer, cervical cancer, ureteral cancer, osteosarcoma, neuroblastoma, fibrosarcoma, rhabdomyosarcoma, astrocytoma, neuroblastoma and glioma, but not limited thereto. Preferably, the cancer that can be treated with the pharmaceutical composition or preparation according to the present invention can be selected from the group consisting of gastric cancer, colorectal cancer, breast cancer, lung cancer and kidney cancer, but not limited thereto.

Autoimmune diseases that can be treated using the pharmaceutical composition or preparation according to the invention include rheumatoid arthritis, asthma, psoriasis, multiple sclerosis, allergic rhinitis, Crohn's disease, ulcerative colitis , systemic lupus erythematosus, type 1 diabetes, inflammatory bowel disease (inflammatory bowel disease, IBD) and atopic dermatitis, but not limited thereto.

The present invention also provides a method for treating diseases, comprising administering the pharmaceutical composition or preparation according to the present invention to a subject in need of treatment, and the present invention further provides a use of the pharmaceutical composition or preparation according to the present invention for treating diseases.

Unless defined otherwise, technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. In addition, repeated descriptions of technical configurations and operations identical to those in the known art will be omitted.

Hereinafter, the present invention will be further explained in detail with reference to examples. These examples are provided for illustrative purposes only, and it is obvious to those having ordinary skill in the art that these examples should not be construed as limiting the scope of the present invention.

[example]

[Example 1, formulation development]

As shown in Table 6, four trastuzumab formulations for subcutaneous injection were prepared. Formulations 1 to 4 typically contained 120 mg/ml of trastuzumab and consisted of 20 mM histidine/histidine-HCl (pH 5.5), 210 mM trehalose, 10 mM methionine, and the pH20 variant. The difference among formulations 1 to 4 is the concentration of nonionic surfactant, where formulation 1: 0% polysorbate 20, formulation 2: 0.005% polysorbate 20, formulation 3: 0.04% polysorbate 20, formulation 4: 0.1% polysorbate 20.

Table 6, Composition of the formulation Preparation 1 Preparation 2 Preparation 3 Preparation 4 Antibody Trastuzumab (120mg/ml) buffer 20 mM Histidine/Histidine-HCl Stabilizer 1 210 mM Trehalose Stabilizer 2 10 mM methionine Polysorbate 20 0% 0.005% 0.04% 0.1% Hyaluronidase HP46 of SEQ ID NO: 44 (2,000 units/ml)

[Example 2, measurement using a spectrophotometer]

Formulations 1 to 4 were left at 45° C. for 14 days, and changes in protein concentration were analyzed using a spectrophotometer manufactured by Beckman. Each sample was diluted with distilled water so that the concentration of the sample was 0.4 mg/ml, and then the absorbance of the protein at 280 nm was measured using a spectrophotometer. In the stability test under harsh conditions (ie, 14 days at 45°C), there was no significant change in the protein concentration of Formulations 1 to 4. However, the activity of hyaluronidase decreased drastically at 45°C, so in this example, the enzyme activity was not measured (see Figure 6).

[Example 3, Studying the Monomer Ratio of Trastuzumab in Each Formulation Using Particle Size Sizing Chromatography]

For size-exclusion chromatography analysis (size-exclusion chromatography analysis), use the HPLC system provided by Shimadzu Prominence and TSK-gel G3000SWXL (7.8 X 300 mm, 5 microns) and TSK guard column (TSK guard column, 6.0 x 4.0 mm, 7 microns). 0.2 M potassium phosphate (pH 6.2) containing 0.25 M potassium chloride was used as the mobile phase. The analysis was performed by applying isocratic separation mode at a flow rate of 0.5 ml/min for 35 minutes. The sample was diluted with the assay solvent so that the final concentration was 10 mg/ml. After injecting 20 μl of the sample into the HPLC column, the absorbance at 280 nm of the eluate from the column was recorded. The monomer ratio of Trastuzumab in the HPLC chromatogram was calculated and plotted.

Formulations 1 to 4 exhibited a similar pattern of change when subjected to size sizing chromatography analysis in a stability test under harsh conditions (ie, 14 days at 45°C). The main changes were increased high molecular weight (HMW) and low molecular weight (LMW) degradation products, and decreased monomer content (about 1.5%), and there were no significant differences depending on the formulation. In conclusion, the results of particle size sizing chromatography analysis in the stability test under harsh conditions (i.e. at 45°C) are that, depending on the concentration of polysorbate 20 (0-0.1 % (w/v) ), there was no significant difference in stability between formulations (see Figure 1A~Figure 1B).

[Example 4, Measuring the protein aggregation temperature of formulations containing Trastuzumab and HP46]

The denaturation properties of proteins due to heat were analyzed using Dynamic light scattering (DLS). In this experiment, the size change of protein molecules as a function of temperature change is measured and used to calculate the protein aggregation temperature. For the dynamic light scattering analysis, a Zetasizer-nano-ZS instrument and a quartz cuvette (quartz cuvette, ZEN2112) provided by Malvern were used. During the analysis, the temperature was increased from 25°C to 85°C in 1°C intervals, the samples were diluted to 1 mg/ml with the respective formulation buffers, and 150 µL of the samples were added to the cuvette for analysis .

The aggregation temperature of formulation 1 without polysorbate 20 was 74°C, and the aggregation temperature of formulations 2 to 4 was 76°C (see Figure 2A and Figure 2B ).

[Example 5, Weak Cation Exchange Chromatography Measurement of Formulations Containing Trastuzumab and HP46]

For weak cation exchange chromatography analysis (WCX chromatography analysis), use the HPLC system provided by Shimadzu Prominence and use TSKgel CM-STAT column (TSKgel CM-STAT column, 4.6 x 100 mm, 7 microns), TSKgel protection gel CMSTAT ( TSKgel guard gel CMSTAT, inner diameter 3.2 mm x 1.5 cm) and the like as strings. Mobile phase A was 10 mM sodium phosphate (pH 7.5) and mobile phase B was 10 mM sodium phosphate (pH 7.2) containing 0.1 M sodium chloride. Analysis was performed with a linear concentration gradient of 0-30% mobile phase B at a flow rate of 0.8 ml/min for 55 minutes. Dilute the sample with mobile phase A so that the final concentration is 1.0 mg/ml, inject 80 μl of the sample into HPLC, and then record the absorbance of the eluate from the column at 280 nm. The monomer ratio of trastuzumab in the HPLC chromatogram was calculated and plotted.

Formulations 1 to 4 exhibited similar patterns of change when analyzed by weak cation exchange chromatography in a stability test under harsh conditions (ie, 14 days at 45°C). Specific changes include an increase in the relative content of acidic variants (about 30% change in 14 days), a decrease in the relative content of the main peak (about 44% change in 14 days), and an increase in the relative content of basic variants (about 15% change in 14 days), and There were no significant differences by formulation. In conclusion, protein stability based on polysorbate 20 (0-0.1%) was similar in weak cation exchange chromatography analysis in the stability test under harsh conditions (i.e. at 45°C) (please See Figure 3A~Figure 3D).

[Example 6, formulation development]

As shown in Table 7, three trastuzumab formulations for subcutaneous injection were prepared. Formulations 5 to 7 generally contained 120 mg/ml of trastuzumab, 20 mM histidine/histidine-HCl (pH 5.5), 210 mM trehalose, 10 mM methionine, and HP46. The difference among Formulations 5 to 7 is the composition of Stabilizer 3, where Formulation 5: 0.04% Polysorbate 20, Formulation 6: 50 mM Lys-Lys, Formulation 7: Glycine.

Table 7, Composition of the formulation Preparation 5 Preparation 6 Preparation 7 Antibody Trastuzumab (120mg/ml) buffer 20 mM Histidine/Histidine-HCl Stabilizer 1 210 mM Trehalose Stabilizer 2 10 mM methionine Stabilizer 3 0.04% Polysorbate 20 50 mM Lys-Lys 50 mM glycine Hyaluronidase HP46 of SEQ ID NO: 44 (2,000 units/ml)

[Example 7, measured using a spectrophotometer]

Formulations 5 to 7 were left at 45° C. for 14 days, and changes in protein concentration were analyzed using a spectrophotometer manufactured by Beckman. Each sample was diluted with distilled water so that the concentration of the sample was 0.4 mg/ml, and then the absorbance of the protein at 280 nm was measured using a spectrophotometer. In the stability test under harsh conditions (ie, 14 days at 45°C), there was no significant change in the protein concentration of Formulations 5 to 7. However, the activity of hyaluronidase decreased drastically at 45°C, so in this example, the enzyme activity was not measured (see Figure 6).

[Example 8, Studying the Monomer Ratio of Trastuzumab in Each Formulation Using Particle Size Sizing Chromatography]

For particle size sizing chromatography analysis, an HPLC system provided by Shimadzu Prominence was used and TSK-gel G3000SWXL (7.8 x 300 mm, 5 microns) and a TSK guard column (6.0 x 4.0 mm, 7 microns) were used as columns. 0.2 M potassium phosphate (pH 6.2) containing 0.25 M potassium chloride was used as the mobile phase. The isocratic separation mode was applied at a flow rate of 0.5 ml/min for 35 minutes. The sample was diluted with the assay solvent so that the final concentration was 10 mg/ml, and the absorbance at 280 nm was measured after injecting 20 μl of the sample into the HPLC column. The monomer ratio of trastuzumab in the HPLC chromatogram was calculated and plotted.

Formulations 5 to 7 exhibited a similar pattern of change when subjected to particle size sizing chromatography analysis in a stability test under harsh conditions (ie, 14 days at 45°C). The main changes are the increase of high molecular weight (HMW) and low molecular weight (LMW) impurities, and the decrease of monomer content (about 1.5%), and there is no significant difference according to the formulation. In conclusion, in the stability test under harsh conditions (i.e. at 45°C), the results of particle size sizing chromatography analysis showed that in 0.04% polysorbate 20, 50 mM Lys-Lys and 50 mM glycerol Amino acid formulations exhibited similar protein stability (see Figure 4).

[Example 9, weak cation exchange chromatography analysis of formulations comprising Trastuzumab and HP46]

For weak cation exchange chromatography analysis, use an HPLC system provided by Shimadzu Prominence and use TSKgel CM-STAT (4.6 x 100 mm, 7 µm), TSKgel protection gel CMSTAT (inner diameter 3.2 mm x 1.5 cm) and the like as tubes column. Mobile phase A was 10 mM sodium phosphate (pH 7.5) and mobile phase B was 10 mM sodium phosphate (pH 7.2) containing 0.1 M sodium chloride. Analysis was performed by separation mode using a linear concentration gradient of 0-30% mobile phase B at a flow rate of 0.8 ml/min for 55 min. Samples were diluted using mobile phase A to a final concentration of 1.0 mg/ml, 80 microliters of the sample were injected into the HPLC, and the absorbance at 280 nm was recorded. The monomer ratio of trastuzumab in the HPLC chromatogram was calculated and plotted.

Formulations 5 to 7 exhibited a similar pattern of change when analyzed by weak cation exchange chromatography in a stability test under harsh conditions (ie, 14 days at 45°C). Specific changes include an increase in the relative content of acidic variants (about 30% change in 14 days), a decrease in the relative content of the main peak (about 44% change in 14 days), and an increase in the relative content of basic variants (about 15% change in 14 days), and There were no significant differences by formulation. In conclusion, weak cation-exchange chromatography analysis in a stability test under harsh conditions (i.e. at 45°C) showed that in 0.04% polysorbate 20, 50 mM Lys-Lys and 50 mM glycine Acid formulations exhibited similar protein stability (see Figures 5A-5C).

[Example 10, Stability evaluation of HP46 at temperatures of 40°C and 45°C in subcutaneous formulations of Trastuzumab and HP46]

To assess the stability of HP46 in a subcutaneous formulation of trastuzumab, trastuzumab (120 mg/ml) was mixed with PH20 (2000 units/ml). At this time, the buffer used contained 20 mM histidine (pH 5.5), 210 mM trehalose, 10 mM methionine, and 0.04% polysorbate 20. The enzyme activity of the samples in the control group was measured on day 0, while the samples in the experimental group were placed at 40°C or 45°C for 1 day, and then the enzyme activity of each sample was measured.

Each Heaiping subcutaneous injection preparation (trastuzumab + HW2 and trastuzumab + HP46) was placed at 40°C for 1 day, and then the activity of hyaluronidase was measured. As a result, each condition showed 51%, 47% and 94% respectively. activity, which means that HP46 has the best thermal stability at 40°C (see Figure 6A~Figure 6B). In addition, He Aiping subcutaneous injection preparations (trastuzumab + HW2 and trastuzumab + HP46) were placed at 45°C for 1 day, and then the activity of hyaluronidase was measured. As a result, He Aiping subcutaneous injection preparations and trastuzumab + HW2 did not have Enzyme activity, but the enzyme activity of trastuzumab + HP46 still exists (see Figure 6~Figure 6B).

[Example 11, formulation development]

As shown in Table 8, three formulations of trastuzumab for subcutaneous injection were prepared. Formulations 8 to 10 generally contained trastuzumab at 120 mg/ml, 20 mM histidine/histidine-HCl (pH 5.5), 210 mM trehalose, 10 mM methionine, and the pH20 variant. The difference among formulations 8 to 10 is the concentration of nonionic surfactant, where formulation 8: 0% polysorbate 20, formulation 9: 0.005% polysorbate 20, formulation 10: 0.04% polysorbate 20.

Table 8, Composition of the formulations Preparation 8 Preparation 9 Preparation 10 Antibody Trastuzumab (120mg/ml) Polysorbate 20 0% 0.005% 0.04% buffer 20 mM Histidine/Histidine-HCl Stabilizer 1 210 mM trehalose Stabilizer 2 10 mM methionine pH 5.5 Hyaluronidase HP46 of SEQ ID NO: 44 (2,000 units/ml)

[Example 12, measurement using a spectrophotometer]

Formulations 8 to 10 were left at 40° C. for 14 days, and changes in protein concentration were analyzed using a spectrophotometer manufactured by Beckman. Each sample was diluted with distilled water so that the concentration of the sample was 0.4 mg/ml, and then the absorbance of the protein at 280 nm was measured using a spectrophotometer. In the stability test under harsh conditions (ie, 14 days at 40°C), there was no significant change in protein concentration from Formulation 8 to Formulation 10.

[Example 13, Studying the Monomer Ratio of Trastuzumab in Each Formulation Using Size Sizing Chromatography]

For particle size sizing chromatography analysis, an HPLC system provided by Shimadzu Prominence was used and TSK-gel G3000SWXL (7.8 x 300 mm, 5 microns) and a TSK guard column (6.0 x 4.0 mm, 7 microns) were used as columns. 0.2 M potassium phosphate (pH 6.2) containing 0.25 M potassium chloride was used as the mobile phase. The analysis was performed by applying isocratic separation mode at a flow rate of 0.5 ml/min for 35 minutes. The sample was diluted with the assay solvent so that the final concentration was 10 mg/ml, and the absorbance at 280 nm was measured after injecting 20 μl of the sample into the HPLC column. The monomer ratio of Trastuzumab in the HPLC chromatogram was calculated and plotted.

Formulations 8 to 10 exhibited a similar pattern of change when subjected to particle size sizing chromatography analysis in a stability test under harsh conditions (ie, 14 days at 40°C). The main changes were the increase of high molecular weight (HMW) and low molecular weight (LMW) degradation products, and the decrease of monomer content (approximately less than 1.0%), and there were no significant differences depending on the formulation. In conclusion, the particle size sizing chromatography analysis in the stability test under harsh conditions (i.e. at 40°C) showed that there was a significant difference between formulations depending on the concentration of polysorbate 20 (0-0.04%). There was no significant difference in stability (see Figure 7).

[Example 14, Measuring the protein aggregation temperature of formulations containing Trastuzumab and HP46]

In the field of protein medicine, dynamic light scattering (DLS) is used to analyze the denaturation property of protein caused by heat. In this experiment, the size change of protein molecules as a function of temperature change is measured and used to calculate the protein aggregation temperature. For dynamic light scattering analysis, a Zetasizer-nano-ZS instrument and a quartz optical cell (ZEN2112) provided by Malvern were used. During the analysis, the temperature was increased from 25°C to 85°C in 1°C intervals, the samples were diluted to 1 mg/ml with the respective formulation buffers, and 150 µL of the samples were added to the cuvette for analysis .

Formulation 8, which did not contain polysorbate 20, had an aggregation temperature of 78.3°C, formulation 9 exhibited an aggregation temperature of 77.3°C, and formulation 10 exhibited an aggregation temperature of 77.7°C. In Example 13, even if polysorbate 20 was not included, there was no change in the monomer ratio of the protein. Comparing the results of the case where polysorbate 20 was included and the case where polysorbate 20 was not included, it was confirmed that the protein There is no difference in aggregation between. These results indicate that for the subcutaneous formulation of trastuzumab, it is not necessary to minimize the amount of polysorbate 20 (see Figures 8A-8B).

[Example 15, Weak Cation Exchange Chromatography Analysis of Formulations Containing Trastuzumab and HP46]

For weak cation exchange chromatography analysis, use HPLC system provided by Shimadzu Prominence and use TSKgel CM-STAT column (4.6 x 100 mm, 7 µm), TSKgel protection gel CMSTAT (3.2 mm inner diameter x 1.5 cm) and the like as a column. Mobile phase A was 10 mM sodium phosphate (pH 7.5) and mobile phase B was 10 mM sodium phosphate (pH 7.2) containing 0.1 M sodium chloride. Analysis was performed with a linear concentration gradient of 0-30% mobile phase B at a flow rate of 0.8 ml/min for 55 minutes. Dilute the sample with mobile phase A so that the final concentration is 1.0 mg/ml, inject 80 μl of the sample into HPLC, and then record the absorbance of the eluate from the column at 280 nm. The monomer ratio of trastuzumab in the HPLC chromatogram was calculated and plotted.

Formulations 8 to 10 exhibited a similar pattern of change when analyzed by weak cation exchange chromatography in a stability test under harsh conditions (ie, 14 days at 40°C). Specific changes include an increase in the relative content of acidic variants (about 10% change in 14 days), a decrease in the relative content of the main peak (about 40% change in 14 days), and an increase in the relative content of basic variants (about 300% change in 14 days), and There were no significant differences by formulation. In conclusion, protein stability based on polysorbate 20 (0-0.04%) was similar in weak cation exchange chromatography analysis in the stability test under harsh conditions (i.e. at 40°C) (please See Figure 9A~Figure 9D).

[Example 16, Measuring Enzyme Activity of Preparations Containing Trastuzumab and HP46]

The Turbidimetric assay used to measure enzyme activity is a method that uses the absorbance value to measure the degree of aggregation formed by the combination of residual hyaluronic acid and acidified albumin (BSA), when hyaluronic acid is hydrolyzed by PH20 When , the amount bound to albumin decreases, resulting in a decrease in absorbance. Dilute BTH (Sigma) as a standard to 1 unit/ml, 2 units/ml, 5 units/ml, 7.5 units/ml, 10 units/ml, 15 units/ml, 20 units/ml, 30 units/ml , 50 units/ml and 60 units/ml, and prepared in each tube (tube). The purified PH20 variant samples were diluted to 100X, 300X, 600X, 1200X and 2400X, and prepared in each tube. In clean tubes, a hyaluronidase solution having a concentration of 3 mg/ml was diluted 10 times to a concentration of 0.3 mg/ml so that the volume of each tube became 180 microliters. 60 μl of the sample containing hyaluronidase was added to the diluted hyaluronic acid solution and mixed therewith, and reacted at 37° C. for 45 minutes. After the reaction was completed, 50 microliters of the reacted enzyme and 250 microliters of acidic albumin solution were added to each well of the 96-well plate and shaken for 10 minutes, then the absorbance at 600 nm was measured using a spectrophotometer value.

As a result of the activity analysis in the stability test under harsh conditions (i.e. 14 days at 40°C), it was confirmed that the higher the concentration of polysorbate 20, the more the activity decreased over time (see Figure 10 ).

[Example 17, formulation development]

As shown in Table 9, three trastuzumab formulations for subcutaneous injection were prepared. Formulations 11 to 13 generally contained 120 mg/ml of trastuzumab, 20 mM histidine/histidine-HCl (pH 5.5), 210 mM trehalose, 10 mM methionine, and the pH20 variant. The difference among formulations 11 to 13 is the concentration of nonionic surfactant, where formulation 11: 0% polysorbate 80, formulation 12: 0.005% polysorbate 80, formulation 13: 0.04% polysorbate 80.

Table 9. Composition of the formulations Preparation 11 Preparation 12 Preparation 13 Antibody Trastuzumab (120 mg/mL) Polysorbate 80 0% 0.005% 0.04% buffer 20 mM Histidine/Histidine-HCl Stabilizer 1 210 mM Trehalose Stabilizer 2 10 mM methionine pH 5.5 Hyaluronidase HP46 of SEQ ID NO: 44 (2,000 units/ml)

Formulations 11 to 13 exhibited a similar pattern of change when subjected to size sizing chromatography analysis in a stability test under harsh conditions (ie, 14 days at 40°C). The main changes were the increase of high molecular weight (HMW) and low molecular weight (LMW) degradation products, and the decrease of monomer content (approximately less than 1.0%), and there were no significant differences depending on the formulation. In conclusion, the particle size sizing chromatography analysis in the stability test under harsh conditions (i.e. at 40°C) showed that there was a significant difference between formulations depending on the concentration of polysorbate 80 (0-0.04%). There was no significant difference in stability (see Figure 11).

[Example 18, Weak Cation Exchange Chromatography Analysis of Formulations Containing Trastuzumab and HP46]

For weak cation exchange chromatography analysis, use HPLC system provided by Shimadzu Prominence and use TSKgel CM-STAT column (4.6 x 100 mm, 7 µm), TSKgel protection gel CMSTAT (3.2 mm inner diameter x 1.5 cm) and the like as a column. Mobile phase A was 10 mM sodium phosphate (pH 7.5) and mobile phase B was 10 mM sodium phosphate (pH 7.2) containing 0.1 M sodium chloride. Analysis was performed with a linear concentration gradient of 0-30% mobile phase B at a flow rate of 0.8 ml/min for 55 minutes. Dilute the sample with mobile phase A so that the final concentration is 1.0 mg/ml, inject 80 μl of the sample into HPLC, and then record the absorbance of the eluate from the column at 280 nm. The monomer ratio of trastuzumab in the HPLC chromatogram was calculated and plotted.

Formulations 11 to 13 exhibited a similar pattern of change when analyzed by weak cation exchange chromatography in a stability test under harsh conditions (ie, 14 days at 40°C). Specific changes include an increase in the relative content of acidic variants (about 10% change in 14 days), a decrease in the relative content of the main peak (about 40% change in 14 days), and an increase in the relative content of basic variants (about 300% change in 14 days), and There were no significant differences by formulation. In conclusion, protein stability based on polysorbate 80 (0-0.04%) was similar in weak cation exchange chromatography analysis in the stability test under harsh conditions (i.e. at 40°C) (please See Figure 12A ~ Figure 12D).

[Example 19, Measuring Enzyme Activity of Preparations Containing Trastuzumab and HP46]

The Turbidimetric assay used to measure enzyme activity is a method that uses the absorbance value to measure the degree of aggregation formed by the combination of residual hyaluronic acid and acidified albumin (BSA), when hyaluronic acid is hydrolyzed by PH20 When , the amount bound to albumin decreases, resulting in a decrease in absorbance. Dilute BTH (Sigma) as a standard to 1 unit/ml, 2 units/ml, 5 units/ml, 7.5 units/ml, 10 units/ml, 15 units/ml, 20 units/ml, 30 units/ml, 50 units/ml and 60 units/ml, and prepared in each tube (tube). Purified protein samples were diluted to 100X, 300X, 600X, 1200X and 2400X with enzyme dilution buffer (20 mM Tris·HCl, pH 7.0, 77 mM NaCl, 0.01% (w/v) bovine serum albumin), And prepared in each tube. In clean tubes, a hyaluronidase solution having a concentration of 3 mg/ml was diluted 10 times to a concentration of 0.3 mg/ml so that the volume of each tube became 180 microliters. 60 μl of the sample containing hyaluronidase was added to the diluted hyaluronic acid solution and mixed therewith, and reacted at 37° C. for 45 minutes. After the reaction was completed, 50 microliters of the reacted enzyme and 250 microliters of acidic albumin solution were added to each well of the 96-well plate and shaken for 10 minutes, then the absorbance at 600 nm was measured using a spectrophotometer value.

As a result of the activity analysis in the stability test under severe conditions (i.e. 14 days at 40°C), it was confirmed that the higher the concentration of polysorbate 80, the more the activity decreased over time (see Figure 13 ).

[Example 20, formulation development]

As shown in Table 10, three formulations of rituximab were prepared. Formulations 14 to 16 generally contained rituximab at 120 mg/ml, 20 mM histidine/histidine-HCl (pH 5.5), 210 mM trehalose, 10 mM methionine, and the pH20 variant. The difference among formulations 14 to 16 is the concentration of nonionic surfactant, where formulation 14: 0% polysorbate 80, formulation 15: 0.005% polysorbate 80, formulation 16: 0.06% polysorbate 80.

Table 10, composition of preparations Preparation 14 Preparation 15 Preparation 16 Rituximab 120 mg/ml (± 10) PS 80 0% 0.005% 0.06% buffer 20 mM Histidine/Histidine-HCl Stabilizer 1 210 mM trehalose Stabilizer 2 10 mM methionine pH 5.5 Hyaluronidase HP46 of SEQ ID NO: 44 (2,000 units/ml)

Formulations 14 to 16 exhibited a similar pattern of change when particle size sizing chromatography was performed in the stability test under harsh conditions (ie, 7 days at 40°C). The main changes were the increase of high molecular weight (HMW) and low molecular weight (LMW) degradation products, and the decrease of monomer content (approximately less than 1.0%), and there were no significant differences depending on the formulation. In conclusion, the particle size sizing chromatography analysis in the stability test under harsh conditions (i.e. at 40°C) showed that there was a significant difference between formulations depending on the concentration of polysorbate 80 (0-0.06%). There was no significant difference in the stability of (see Figure 14).

[Example 21, Measuring Enzyme Activity of Preparations Containing Rituximab and HP46]

The Turbidimetric assay used to measure enzyme activity is a method that uses the absorbance value to measure the degree of aggregation formed by the combination of residual hyaluronic acid and acidified albumin (BSA), when hyaluronic acid is hydrolyzed by PH20 When , the amount bound to albumin decreases, resulting in a decrease in absorbance. Dilute BTH (Sigma) as a standard to 1 unit/ml, 2 units/ml, 5 units/ml, 7.5 units/ml, 10 units/ml, 15 units/ml, 20 units/ml, 30 units/ml , 50 units/ml and 60 units/ml, and prepared in each tube (tube). Purified protein samples were diluted to 100X, 300X, 600X, 1200X and 2400X with enzyme dilution buffer (20 mM Tris·HCl, pH 7.0, 77 mM NaCl, 0.01% (w/v) bovine serum albumin), And prepared in each tube. In clean tubes, a hyaluronidase solution having a concentration of 3 mg/ml was diluted 10 times to a concentration of 0.3 mg/ml so that the volume of each tube became 180 microliters. 60 μl of the sample containing hyaluronidase was added to the diluted hyaluronic acid solution and mixed therewith, and reacted at 37° C. for 45 minutes. After the reaction was completed, 50 microliters of the reacted enzyme and 250 microliters of acidic albumin solution were added to each well of the 96-well plate and shaken for 10 minutes, then the absorbance at 600 nm was measured using a spectrophotometer value.

As a result of the activity analysis in the stability test under severe conditions (i.e. 7 days at 40°C), it was confirmed that the higher the concentration of polysorbate 80, the more the activity decreased over time (see Figure 15 ).

[Example 22, Measuring Enzyme Activity of Commercially Available Formulations Not Containing Polysorbate]

As shown in Table 11, two commercially available rituximab formulations were prepared. Formulation 17 is a commercially available buffer for subcutaneous injection formulation and formulation 18 is a commercially available buffer for intravenous injection formulation. Formulations 17 and 18 included the PH20 variant and included rituximab at 120 mg/ml and rituximab at 100 mg/ml, respectively, but did not include polysorbate 80, unlike the commercially available product.

Table 11, Composition of the formulations Preparation 17 Preparation 18 Rituximab 120 mg/ml 100 mg/ml buffer 20 mM Histidine/Histidine-HCl 25 mM sodium citrate Stabilizer 1 210 mM trehalose 145 mM NaCl Stabilizer 2 10 mM methionine 10 mM methionine pH 5.5 6.5 Hyaluronidase HP46 of SEQ ID NO: 44 (2,000 units/ml)

The turbidity method used to measure enzyme activity is a method that measures the degree of aggregation formed by the binding of residual hyaluronic acid to acidified albumin (BSA) by absorbance. When hyaluronic acid is hydrolyzed by pH20, it binds to The amount of albumin decreases, resulting in a lower absorbency. Dilute BTH (Sigma) as a standard to 1 unit/ml, 2 units/ml, 5 units/ml, 7.5 units/ml, 10 units/ml, 15 units/ml, 20 units/ml, 30 units/ml , 50 units/ml and 60 units/ml, and prepared in separate tubes. Purified protein samples were diluted to 100X, 300X, 600X, 1200X and 2400X with enzyme dilution buffer (20 mM Tris·HCl, pH 7.0, 77 mM NaCl, 0.01% (w/v) bovine serum albumin), And prepared in each tube. In clean tubes, a hyaluronidase solution having a concentration of 3 mg/ml was diluted 10 times to a concentration of 0.3 mg/ml so that the volume of each tube became 180 microliters. 60 μl of the sample containing hyaluronidase was added to the diluted hyaluronic acid solution and mixed therewith, and reacted at 37° C. for 45 minutes. After the reaction was completed, 50 microliters of the reacted enzyme and 250 microliters of acidic albumin solution were added to each well of the 96-well plate and shaken for 10 minutes, then the absorbance at 600 nm was measured using a spectrophotometer value.

As a result of the activity analysis in the stability test under severe conditions (i.e., 6 days at 40 ° C), it was confirmed that even if polysorbate 80 was not included in the formulation, high activity could still be maintained, especially, the formulation 18 maintains high activity (see Figure 16).

[Example 23: Preparation development]

As shown in Table 12, four formulations of pembrolizumab were prepared. Formulation 19, Formulation 20 and Formulation 21 typically contained 25 mg/ml of pembrolizumab, 10 mM histidine (pH 5.5), 7% sucrose, 10 mM methionine, and the pH20 variant. The difference among formulations 19 to 21 is the concentration of non-ionic surfactant, where formulation 19: 0% polysorbate 80, formulation 20: 0.005% polysorbate 80, formulation 21: 0.02% polysorbate 80. Formulation 22 contained 25 mg/ml of pembrolizumab and consisted of 10 mM histidine (pH 5.5), 210 mM trehalose, 10 mM methionine, 0.02% polysorbate 80, and the pH20 variant.

Table 12. Composition of the formulations Preparation 19 Preparation 20 Preparation 21 Preparation 22 Antibody Pembrolizumab (25 mg/mL) buffer 10 mM histidine (pH 5.5) Stabilizer 1 7% sucrose 7% sucrose 7% sucrose 210 mM trehalose Stabilizer 2 10 mM Methionine 10 mM Methionine 10 mM Methionine 10 mM Methionine Polysorbate 80 0% 0.005% 0.02% 0.02% Hyaluronidase HP46 of SEQ ID NO: 44 (2,000 units/ml)

[Example 24, Measurement using a spectrophotometer]

Formulations 19 to 22 were left at 40° C. for 7 days, and changes in protein concentration were analyzed using a spectrophotometer manufactured by Beckman. Each sample was diluted with distilled water so that the concentration of the sample was 0.4 mg/ml, and then the absorbance of the protein at 280 nm was measured using a spectrophotometer.

In the stability test under harsh conditions (ie, 7 days at 40°C), there was no significant change in protein concentration from Formulation 19 to Formulation 22.

[Example 25, Investigation of the monomer ratio of pembrolizumab in various formulations using particle size sizing chromatography]

For particle size sizing chromatography analysis, an HPLC system provided by Shimadzu Prominence was used and TSK-gel G3000SWXL (7.8 x 300 mm, 5 microns) and a TSK guard column (6.0 x 4.0 mm, 7 microns) were used as columns. 0.2 M potassium phosphate (pH 6.2) containing 0.25 M potassium chloride was used as the mobile phase. The analysis was performed by applying isocratic separation mode at a flow rate of 0.5 ml/min for 35 minutes. The sample was diluted with the assay solvent so that the final concentration was 10 mg/ml, and the absorbance at 280 nm was measured after injecting 20 μl of the sample into the HPLC column. The monomer ratio of pembrolizumab in the HPLC chromatogram was calculated and plotted.

Formulations 19 to 22 exhibited a similar pattern of change when particle size sizing chromatography was performed in a stability test under harsh conditions (ie, 7 days at 40°C). There were no significant differences depending on the formulation in the high molecular weight (HMW) and low molecular weight (LMW) degradation products. In conclusion, Formulations 19 to 22 did not exhibit significant differences as a result of particle size sizing chromatography analysis in the stability test under harsh conditions (i.e. at 40°C) and were not dependent on sugar There are differences depending on the type (see Figure 17). These results are consistent with those in the case of trastuzumab and rituximab according to the previous example.

[Example 26, Measuring the Enzyme Activity of a Preparation Containing Pembrolizumab and HP46]

The turbidity method used to measure enzyme activity is a method that measures the degree of aggregation formed by the binding of residual hyaluronic acid to acidified albumin (BSA) by absorbance. When hyaluronic acid is hydrolyzed by pH20, it binds to The amount of albumin decreases, resulting in a lower absorbency. Dilute BTH (Sigma) as a standard to 1 unit/ml, 2 units/ml, 5 units/ml, 7.5 units/ml, 10 units/ml, 15 units/ml, 20 units/ml, 30 units/ml , 50 units/ml and 60 units/ml, and prepared in each tube (tube). Purified protein samples were diluted to 100X, 300X, 600X, 1200X and 2400X with enzyme dilution buffer (20 mM Tris·HCl, pH 7.0, 77 mM NaCl, 0.01% (w/v) bovine serum albumin), And prepared in each tube. In clean tubes, a hyaluronidase solution having a concentration of 3 mg/ml was diluted 10 times to a concentration of 0.3 mg/ml so that the volume of each tube became 180 microliters. Add 60 microliters of the enzyme-containing sample to the diluted hyaluronic acid solution, mix it, and react at 37°C for 45 minutes. After the reaction was completed, 50 microliters of the reacted enzyme and 250 microliters of acidic albumin solution were added to each well of the 96-well plate and shaken for 10 minutes, then the absorbance at 600 nm was measured using a spectrophotometer value.

As a result of the activity analysis in the stability test under severe conditions (ie, 7 days at 40° C.), it was confirmed that the decrease in activity was large with the increase in the concentration of polysorbate 80. It was also confirmed that when the same amount of polysorbate 80 was included, the decreased activity in the trehalose-containing formulation was smaller than that in the sucrose-containing formulation (see FIG. 18 ).

[Example 27, pH-activity graph of HP46 and wild-type HW2]

For experiments to confirm the pH-activity profiles of HP46 and wild-type HW2, a microturbidimetric assay method was used. For each pH value, a hyaluronic acid buffer for dissolving hyaluronic acid as a substrate and an enzyme buffer for diluting the enzyme were prepared.

A total of three 96-well plates were prepared for the reaction between the enzyme and the substrate, labeled as A, B, and C, and the experiment was performed.

Use 20 mM acetic acid and 70 mM sodium chloride to prepare hyaluronic acid buffer at pH 4.0, 4.5 or 5.0, use 20 mM sodium phosphate and 70 mM sodium chloride to prepare hyaluronic acid at pH 5.5, 6.0, 6.5, 7.0 or 8.0 solution. Dissolve 20 mg of hyaluronic acid in 10 ml of each prepared hyaluronic acid buffer to prepare the final hyaluronic acid substrate solution, and then dilute it with each hyaluronic acid buffer prepared according to pH to prepare a concentration of 0.1 mg/ml , 0.25 mg/ml, 0.45 mg/ml or 0.7 mg/ml of 500 μl of the final solution, 100 μl of each solution was dispensed into each well of the 96-well plate labeled A. Use the hyaluronic acid buffer prepared and diluted according to the concentration as a calibration curve to measure the concentration of hyaluronic acid.

Use 20 mM acetic acid, 0.01% (w/v) BSA, and 70 mM sodium chloride to prepare enzyme buffers at pH 4.0, 4.5, or 5.0, use 20 mM sodium phosphate, 0.01% (w/v) BSA, and 70 mM Sodium chloride was used to prepare enzyme buffers at pH 5.5, 6.0, 6.5, 7.0 or 8.0.

HP46 and wild-type HW2 enzymes were prepared at 10 units/ml with enzyme buffer prepared according to the pH value, and 50 microliters of the final solution was distributed to each well of the 96-well plate marked as B.

Transfer 50 microliters of the sample from each well of the 96-well plate marked A to each well of the 96-well plate marked B, and then react at 37°C, shaking the incubator for 45 minutes. 15 minutes before the completion of the reaction, distribute 200 microliters of acidic albumin solution into each well of the 96-well plate marked C, and when the enzyme substrate reaction is complete, dispense 40 microliters of the sample from the well marked B Each well of the 96-well plate was transferred to each well of the 96-well plate marked C, followed by reaction for 20 minutes. After 20 minutes, measure the absorbance at 600 nm, calculate the amount of hyaluronic acid remaining after the enzyme substrate reaction, and complete the enzyme activity diagram according to the pH value (see Figure 19).

[Example 28, Pharmacokinetics of Heaiping Subcutaneous Injection Formulation with Trastuzumab and HP46 in Sprague-Dawley Rats]

In order to test whether the subcutaneous injection preparations of trastuzumab and HP46 exhibit the same pharmacokinetic properties as the He Aiping subcutaneous injection preparations, 9-week-old Schwartz rats were used for the experiment. The dosage of He Aiping and trastuzumab was 18 mg/kg (mouse body weight), the amount of rHuPH20 contained in He Aiping's subcutaneous injection preparation was 100 U, and the amount of HP46 was also 100 U. In the pharmacokinetic test, trastuzumab and HP46 exhibited the same area under the curve (AUC) as the Heaiping subcutaneous injection preparation (see Figure 20).

The pharmaceutical composition according to the present invention can be used for subcutaneous injection and is quite stable. The activity of the PH20 variant and the drug can be maintained for a long time. The drug is preferably an antibody drug or an analog. Therefore, the pharmaceutical composition not only helps to reduce the cost of producing hypodermic preparations, but also reduces medical costs, which is quite effective in terms of patient convenience.

Although the preferred embodiment of the present invention has been disclosed for illustration, those skilled in the art should understand that various modifications, additions and substitutions can be made without departing from the scope and spirit of the present invention as disclosed in the claims .

〔references〕 Bookbinder, L.H., Hofer, A., Haller, M.F., Zepeda, M.L., Keller, G.A., Lim, J.E., Edgington, T.S., Shepard, H.M., Patton, J.S., and Frost, G.I. (2006). A recombinant human enzyme for Enhanced interstitial transport of therapeutics. J Control Release 114, 230-241. Borders jr., C.L. and Raftery, A. (1968) Purification and Partial Characterization of Testicular Hyaluronidase. J Biol Chem 243, 3756-3762 Chao, K.L., Muthukumar, L., and Herzberg, O. (2007). Structure of human hyaluronidase-1, a hyaluronan hydrolyzing enzyme involved in tumor growth and angiogenesis. Biochemistry 46, 6911-6920. Chen, K.J., Sabrina, S., El-Safory, N.S., Lee, G.C., and Lee, C.K. (2016) Constitutive expression of recombinant human hyaluronidase PH20 by Pichia pastoris. J Biosci Bioeng. 122, 673-678 Frost, G.I. (2007). Recombinant human hyaluronidase (rHuPH20): an enabling platform for subcutaneous drug and fluid administration. Expert Opin Drug Deliv 4, 427-440 Hofinger, E.S., Bernhardt, G., and Buschauer, A. (2007) Kinetics of Hyal-1 and PH-20 hyaluronidases: comparison of minimal substrates and analysis of the transglycosylation reaction. Glycobiology 17, 963-971 Kreidieh, F.Y., Moukadem, H.A., and Saghir, N.S.E. (2016) Overview, prevention and management of chemotherapy extravasation. World J Clin Oncol 7, 87-97. Thomas, J.R., Yocum, R.C., Haller, M.F., and Flament J. (2009) The INFUSE-Morphine IIB Study: Use of Recombinant Human Hyaluronidase (rHuPH20) to Enhance the Absorption of Subcutaneous Morphine in Healthy Volunteers. J Pain Symptom Manag 38 , 673-682

none sequence listing ＜110＞ South Korean company Artegen Co., Ltd. <120> Pharmaceutical composition comprising human hyaluronidase PH20 variant and drug for subcutaneous injection ＜130＞ PP-B2376 ＜160＞ 51 ＜170＞ PatentIn version 3. 5 ＜210＞ 1 ＜211＞ 509 ＜212＞ PRT ＜213＞ Artificial sequence <220> ＜223＞ Synthetic sequence ＜400＞ 1 Met Gly Val Leu Lys Phe Lys His Ile Phe Phe Arg Ser Phe Val Lys 1 5 10 15 Ser Ser Gly Val Ser Gln Ile Val Phe Thr Phe Leu Leu Ile Pro Cys 20 25 30 Cys Leu Thr Leu Asn Phe Arg Ala Pro Pro Val Ile Pro Asn Val Pro 35 40 45 Phe Leu Trp Ala Trp Asn Ala Pro Ser Glu Phe Cys Leu Gly Lys Phe 50 55 60 Asp Glu Pro Leu Asp Met Ser Leu Phe Ser Phe Ile Gly Ser Pro Arg 65 70 75 80 Ile Asn Ala Thr Gly Gln Gly Val Thr Ile Phe Tyr Val Asp Arg Leu 85 90 95 Gly Tyr Tyr Pro Tyr Ile Asp Ser Ile Thr Gly Val Thr Val Asn Gly 100 105 110 Gly Ile Pro Gln Lys Ile Ser Leu Gln Asp His Leu Asp Lys Ala Lys 115 120 125 Lys Asp Ile Thr Phe Tyr Met Pro Val Asp Asn Leu Gly Met Ala Val 130 135 140 Ile Asp Trp Glu Glu Trp Arg Pro Thr Trp Ala Arg Asn Trp Lys Pro 145 150 155 160 Lys Asp Val Tyr Lys Asn Arg Ser Ile Glu Leu Val Gln Gln Gln Asn 165 170 175 Val Gln Leu Ser Leu Thr Glu Ala Thr Glu Lys Ala Lys Gln Glu Phe 180 185 190 Glu Lys Ala Gly Lys Asp Phe Leu Val Glu Thr Ile Lys Leu Gly Lys 195 200 205 Leu Leu Arg Pro Asn His Leu Trp Gly Tyr Tyr Leu Phe Pro Asp Cys 210 215 220 Tyr Asn His His Tyr Lys Lys Pro Gly Tyr Asn Gly Ser Cys Phe Asn 225 230 235 240 Val Glu Ile Lys Arg Asn Asp Asp Leu Ser Trp Leu Trp Asn Glu Ser 245 250 255 Thr Ala Leu Tyr Pro Ser Ile Tyr Leu Asn Thr Gln Gln Ser Pro Val 260 265 270 Ala Ala Thr Leu Tyr Val Arg Asn Arg Val Arg Glu Ala Ile Arg Val 275 280 285 Ser Lys Ile Pro Asp Ala Lys Ser Pro Leu Pro Val Phe Ala Tyr Thr 290 295 300 Arg Ile Val Phe Thr Asp Gln Val Leu Lys Phe Leu Ser Gln Asp Glu 305 310 315 320 Leu Val Tyr Thr Phe Gly Glu Thr Val Ala Leu Gly Ala Ser Gly Ile 325 330 335 Val Ile Trp Gly Thr Leu Ser Ile Met Arg Ser Met Lys Ser Cys Leu 340 345 350 Leu Leu Asp Asn Tyr Met Glu Thr Ile Leu Asn Pro Tyr Ile Ile Asn 355 360 365 Val Thr Leu Ala Ala Lys Met Cys Ser Gln Val Leu Cys Gln Glu Gln 370 375 380 Gly Val Cys Ile Arg Lys Asn Trp Asn Ser Ser Asp Tyr Leu His Leu 385 390 395 400 Asn Pro Asp Asn Phe Ala Ile Gln Leu Glu Lys Gly Gly Lys Phe Thr 405 410 415 Val Arg Gly Lys Pro Thr Leu Glu Asp Leu Glu Gln Phe Ser Glu Lys 420 425 430 Phe Tyr Cys Ser Cys Tyr Ser Thr Leu Ser Cys Lys Glu Lys Ala Asp 435 440 445 Val Lys Asp Thr Asp Ala Val Asp Val Cys Ile Ala Asp Gly Val Cys 450 455 460 Ile Asp Ala Phe Leu Lys Pro Pro Met Glu Thr Glu Glu Pro Gln Ile 465 470 475 480 Phe Tyr Asn Ala Ser Pro Ser Thr Leu Ser Ala Thr Met Phe Ile Val 485 490 495 Ser Ile Leu Phe Leu Ile Ile Ser Ser Val Ala Ser Leu 500 505 ＜210＞ 2 ＜211＞ 26 ＜212＞ PRT ＜213＞ Artificial sequence <220> ＜223＞ Synthetic sequence ＜400＞ 2 Met Ala Thr Gly Ser Arg Thr Ser Leu Leu Leu Ala Phe Gly Leu Leu 1 5 10 15 Cys Leu Pro Trp Leu Gln Glu Gly Ser Ala 20 25 ＜210＞ 3 ＜211＞ 18 ＜212＞ PRT ＜213＞ Artificial sequence <220> ＜223＞ Synthetic sequence ＜400＞ 3 Met Lys Trp Val Thr Phe Ile Ser Leu Leu Phe Leu Phe Ser Ser Ser Ala 1 5 10 15 Tyr Ser ＜210＞ 4 ＜211＞ 21 ＜212＞ PRT ＜213＞ Artificial sequence <220> ＜223＞ Synthetic sequence ＜400＞ 4 Met Ala Ala His Leu Leu Pro Ile Cys Ala Leu Phe Leu Thr Leu Leu 1 5 10 15 Asp Met Ala Gln Gly 20 ＜210＞ 5 ＜211＞ 455 ＜212＞ PRT ＜213＞ Artificial sequence <220> ＜223＞ Synthetic sequence ＜400＞ 5 Leu Asn Phe Arg Ala Pro Pro Val Ile Pro Asn Val Pro Phe Leu Trp 1 5 10 15 Ala Trp Asn Ala Pro Ser Glu Phe Cys Leu Gly Lys Phe Asp Glu Pro 20 25 30 Leu Asp Met Ser Leu Phe Ser Phe Ile Gly Ser Pro Arg Ile Asn Ala 35 40 45 Thr Gly Gln Gly Val Thr Ile Phe Tyr Val Asp Arg Leu Gly Tyr Tyr 50 55 60 Pro Tyr Ile Asp Ser Ile Thr Gly Val Thr Val Asn Gly Gly Ile Pro 65 70 75 80 Gln Lys Ile Ser Leu Gln Asp His Leu Asp Lys Ala Lys Lys Lys Asp Ile 85 90 95 Thr Phe Tyr Met Pro Val Asp Asn Leu Gly Met Ala Val Ile Asp Trp 100 105 110 Glu Glu Trp Arg Pro Thr Trp Ala Arg Asn Trp Lys Pro Lys Asp Val 115 120 125 Tyr Lys Asn Arg Ser Ile Glu Leu Val Gln Gln Gln Asn Val Gln Leu 130 135 140 Ser Leu Thr Glu Ala Thr Glu Lys Ala Lys Gln Glu Phe Glu Lys Ala 145 150 155 160 Gly Lys Asp Phe Leu Val Glu Thr Ile Lys Leu Gly Lys Leu Leu Arg 165 170 175 Pro Asn His Leu Trp Gly Tyr Tyr Leu Phe Pro Asp Cys Tyr Asn His 180 185 190 His Tyr Lys Lys Pro Gly Tyr Asn Gly Ser Cys Phe Asn Val Glu Ile 195 200 205 Lys Arg Asn Asp Asp Leu Ser Trp Leu Trp Asn Glu Ser Thr Ala Leu 210 215 220 Tyr Pro Ser Ile Tyr Leu Asn Thr Gln Gln Ser Pro Val Ala Ala Thr 225 230 235 240 Leu Tyr Val Arg Asn Arg Val Arg Glu Ala Ile Arg Val Ser Lys Ile 245 250 255 Pro Asp Ala Lys Ser Pro Leu Pro Val Phe Ala Tyr Thr Arg Ile Val 260 265 270 Phe Thr Asp Gln Val Leu Lys Phe Leu Ser Gln Asp Glu Leu Val Tyr 275 280 285 Thr Phe Gly Glu Thr Val Ala Leu Gly Ala Ser Gly Ile Val Ile Trp 290 295 300 Gly Thr Leu Ser Ile Thr Arg Thr Lys Glu Ser Cys Gln Ala Ile Lys 305 310 315 320 Glu Tyr Met Asp Thr Thr Leu Gly Pro Tyr Ile Ile Asn Val Thr Leu 325 330 335 Ala Ala Lys Met Cys Ser Gln Val Leu Cys Gln Glu Gln Gly Val Cys 340 345 350 Ile Arg Lys Asn Trp Asn Ser Ser Asp Tyr Leu His Leu Asn Pro Asp 355 360 365 Asn Phe Ala Ile Gln Leu Glu Lys Gly Gly Lys Phe Thr Val Arg Gly 370 375 380 Lys Pro Thr Leu Glu Asp Leu Glu Gln Phe Ser Glu Lys Phe Tyr Cys 385 390 395 400 Ser Cys Tyr Ser Thr Leu Ser Cys Lys Glu Lys Ala Asp Val Lys Asp 405 410 415 Thr Asp Ala Val Asp Val Cys Ile Ala Asp Gly Val Cys Ile Asp Ala 420 425 430 Phe Leu Lys Pro Pro Met Glu Thr Glu Glu Pro Gln Ile Phe Tyr Asn 435 440 445 Ala Ser Pro Ser Thr Leu Ser 450 455 ＜210＞ 6 ＜211＞ 455 ＜212＞ PRT ＜213＞ Artificial sequence <220> ＜223＞ Synthetic sequence ＜400＞ 6 Leu Asn Phe Arg Ala Pro Pro Val Ile Pro Asn Val Pro Phe Leu Trp 1 5 10 15 Ala Trp Asn Ala Pro Ser Glu Phe Cys Leu Gly Lys Phe Asp Glu Pro 20 25 30 Leu Asp Met Ser Leu Phe Ser Phe Ile Gly Ser Pro Arg Ile Asn Ala 35 40 45 Thr Gly Gln Gly Val Thr Ile Phe Tyr Val Asp Arg Leu Gly Tyr Tyr 50 55 60 Pro Tyr Ile Asp Ser Ile Thr Gly Val Thr Val Asn Gly Gly Ile Pro 65 70 75 80 Gln Lys Ile Ser Leu Gln Asp His Leu Asp Lys Ala Lys Lys Lys Asp Ile 85 90 95 Thr Phe Tyr Met Pro Val Asp Asn Leu Gly Met Ala Val Ile Asp Trp 100 105 110 Glu Glu Trp Arg Pro Thr Trp Ala Arg Asn Trp Lys Pro Lys Asp Val 115 120 125 Tyr Lys Asn Arg Ser Ile Glu Leu Val Gln Gln Gln Asn Val Gln Leu 130 135 140 Ser Leu Thr Glu Ala Thr Glu Lys Ala Lys Gln Glu Phe Glu Lys Ala 145 150 155 160 Gly Lys Asp Phe Leu Val Glu Thr Ile Lys Leu Gly Lys Leu Leu Arg 165 170 175 Pro Asn His Leu Trp Gly Tyr Tyr Leu Phe Pro Asp Cys Tyr Asn His 180 185 190 His Tyr Lys Lys Pro Gly Tyr Asn Gly Ser Cys Phe Asn Val Glu Ile 195 200 205 Lys Arg Asn Asp Asp Leu Ser Trp Leu Trp Asn Glu Ser Thr Ala Leu 210 215 220 Tyr Pro Ser Ile Tyr Leu Asn Thr Gln Gln Ser Pro Val Ala Ala Thr 225 230 235 240 Leu Tyr Val Arg Asn Arg Val Arg Glu Ala Ile Arg Val Ser Lys Ile 245 250 255 Pro Asp Ala Lys Ser Pro Leu Pro Val Phe Ala Tyr Thr Arg Ile Val 260 265 270 Phe Thr Asp Gln Val Leu Lys Phe Leu Ser Gln Asp Glu Leu Val Tyr 275 280 285 Thr Phe Gly Glu Thr Val Ala Leu Gly Ala Ser Gly Ile Val Ile Trp 290 295 300 Gly Thr Leu Ser Ile Thr Arg Thr Lys Glu Ser Cys Gln Ala Ile Lys 305 310 315 320 Glu Tyr Met Asp Thr Thr Leu Asn Pro Phe Ile Leu Asn Val Thr Ser 325 330 335 Gly Ala Leu Leu Cys Ser Gln Ala Leu Cys Gln Glu Gln Gly Val Cys 340 345 350 Ile Arg Lys Asn Trp Asn Ser Ser Asp Tyr Leu His Leu Asn Pro Asp 355 360 365 Asn Phe Ala Ile Gln Leu Glu Lys Gly Gly Lys Phe Thr Val Arg Gly 370 375 380 Lys Pro Thr Leu Glu Asp Leu Glu Gln Phe Ser Glu Lys Phe Tyr Cys 385 390 395 400 Ser Cys Tyr Ser Thr Leu Ser Cys Lys Glu Lys Ala Asp Val Lys Asp 405 410 415 Thr Asp Ala Val Asp Val Cys Ile Ala Asp Gly Val Cys Ile Asp Ala 420 425 430 Phe Leu Lys Pro Pro Met Glu Thr Glu Glu Pro Gln Ile Phe Tyr Asn 435 440 445 Ala Ser Pro Ser Thr Leu Ser 450 455 ＜210＞ 7 ＜211＞ 455 ＜212＞ PRT ＜213＞ Artificial sequence <220> ＜223＞ Synthetic sequence ＜400＞ 7 Leu Asn Phe Arg Ala Pro Pro Val Ile Pro Asn Val Pro Phe Leu Trp 1 5 10 15 Ala Trp Asn Ala Pro Ser Glu Phe Cys Leu Gly Lys Phe Asp Glu Pro 20 25 30 Leu Asp Met Ser Leu Phe Ser Phe Ile Gly Ser Pro Arg Ile Asn Ala 35 40 45 Thr Gly Gln Gly Val Thr Ile Phe Tyr Val Asp Arg Leu Gly Tyr Tyr 50 55 60 Pro Tyr Ile Asp Ser Ile Thr Gly Val Thr Val Asn Gly Gly Ile Pro 65 70 75 80 Gln Lys Ile Ser Leu Gln Asp His Leu Asp Lys Ala Lys Lys Lys Asp Ile 85 90 95 Thr Phe Tyr Met Pro Val Asp Asn Leu Gly Met Ala Val Ile Asp Trp 100 105 110 Glu Glu Trp Arg Pro Thr Trp Ala Arg Asn Trp Lys Pro Lys Asp Val 115 120 125 Tyr Lys Asn Arg Ser Ile Glu Leu Val Gln Gln Gln Asn Val Gln Leu 130 135 140 Ser Leu Thr Glu Ala Thr Glu Lys Ala Lys Gln Glu Phe Glu Lys Ala 145 150 155 160 Gly Lys Asp Phe Leu Val Glu Thr Ile Lys Leu Gly Lys Leu Leu Arg 165 170 175 Pro Asn His Leu Trp Gly Tyr Tyr Leu Phe Pro Asp Cys Tyr Asn His 180 185 190 His Tyr Lys Lys Pro Gly Tyr Asn Gly Ser Cys Phe Asn Val Glu Ile 195 200 205 Lys Arg Asn Asp Asp Leu Ser Trp Leu Trp Asn Glu Ser Thr Ala Leu 210 215 220 Tyr Pro Ser Ile Tyr Leu Asn Thr Gln Gln Ser Pro Val Ala Ala Thr 225 230 235 240 Leu Tyr Val Arg Asn Arg Val Arg Glu Ala Ile Arg Val Ser Lys Ile 245 250 255 Pro Asp Ala Lys Ser Pro Leu Pro Val Phe Ala Tyr Thr Arg Ile Val 260 265 270 Phe Thr Asp Gln Val Leu Lys Phe Leu Ser Gln Asp Glu Leu Val Tyr 275 280 285 Thr Phe Gly Glu Thr Val Ala Leu Gly Ala Ser Gly Ile Val Ile Trp 290 295 300 Gly Thr Leu Ser Ile Thr Arg Thr Lys Glu Ser Cys Gln Ala Ile Lys 305 310 315 320 Glu Tyr Met Asp Thr Thr Leu Gly Pro Phe Ile Leu Asn Val Thr Ser 325 330 335 Gly Ala Leu Leu Cys Ser Gln Ala Leu Cys Gln Glu Gln Gly Val Cys 340 345 350 Ile Arg Lys Asn Trp Asn Ser Ser Asp Tyr Leu His Leu Asn Pro Asp 355 360 365 Asn Phe Ala Ile Gln Leu Glu Lys Gly Gly Lys Phe Thr Val Arg Gly 370 375 380 Lys Pro Thr Leu Glu Asp Leu Glu Gln Phe Ser Glu Lys Phe Tyr Cys 385 390 395 400 Ser Cys Tyr Ser Thr Leu Ser Cys Lys Glu Lys Ala Asp Val Lys Asp 405 410 415 Thr Asp Ala Val Asp Val Cys Ile Ala Asp Gly Val Cys Ile Asp Ala 420 425 430 Phe Leu Lys Pro Pro Met Glu Thr Glu Glu Pro Gln Ile Phe Tyr Asn 435 440 445 Ala Ser Pro Ser Thr Leu Ser 450 455 ＜210＞ 8 ＜211＞ 455 ＜212＞ PRT ＜213＞ Artificial sequence <220> ＜223＞ Synthetic sequence ＜400＞ 8 Leu Asn Phe Arg Ala Pro Pro Val Ile Pro Asn Val Pro Phe Leu Trp 1 5 10 15 Ala Trp Asn Ala Pro Ser Glu Phe Cys Leu Gly Lys Phe Asp Glu Pro 20 25 30 Leu Asp Met Ser Leu Phe Ser Phe Ile Gly Ser Pro Arg Ile Asn Ala 35 40 45 Thr Gly Gln Gly Val Thr Ile Phe Tyr Val Asp Arg Leu Gly Tyr Tyr 50 55 60 Pro Tyr Ile Asp Ser Ile Thr Gly Val Thr Val Asn Gly Gly Ile Pro 65 70 75 80 Gln Lys Ile Ser Leu Gln Asp His Leu Asp Lys Ala Lys Lys Lys Asp Ile 85 90 95 Thr Phe Tyr Met Pro Val Asp Asn Leu Gly Met Ala Val Ile Asp Trp 100 105 110 Glu Glu Trp Arg Pro Thr Trp Ala Arg Asn Trp Lys Pro Lys Asp Val 115 120 125 Tyr Lys Asn Arg Ser Ile Glu Leu Val Gln Gln Gln Asn Val Gln Leu 130 135 140 Ser Leu Thr Glu Ala Thr Glu Lys Ala Lys Gln Glu Phe Glu Lys Ala 145 150 155 160 Gly Lys Asp Phe Leu Val Glu Thr Ile Lys Leu Gly Lys Leu Leu Arg 165 170 175 Pro Asn His Leu Trp Gly Tyr Tyr Leu Phe Pro Asp Cys Tyr Asn His 180 185 190 His Tyr Lys Lys Pro Gly Tyr Asn Gly Ser Cys Phe Asn Val Glu Ile 195 200 205 Lys Arg Asn Asp Asp Leu Ser Trp Leu Trp Asn Glu Ser Thr Ala Leu 210 215 220 Tyr Pro Ser Ile Tyr Leu Asn Thr Gln Gln Ser Pro Val Ala Ala Thr 225 230 235 240 Leu Tyr Val Arg Asn Arg Val Arg Glu Ala Ile Arg Val Ser Lys Ile 245 250 255 Pro Asp Ala Lys Ser Pro Leu Pro Val Phe Ala Tyr Thr Arg Ile Val 260 265 270 Phe Thr Asp Gln Val Leu Lys Phe Leu Ser Gln Asp Glu Leu Val Tyr 275 280 285 Thr Phe Gly Glu Thr Val Ala Leu Gly Ala Ser Gly Ile Val Ile Trp 290 295 300 Val Ser Trp Glu Asn Thr Arg Thr Lys Glu Ser Cys Gln Ala Ile Lys 305 310 315 320 Glu Tyr Met Asp Thr Thr Leu Gly Pro Tyr Ile Ile Asn Val Thr Leu 325 330 335 Ala Ala Lys Met Cys Ser Gln Val Leu Cys Gln Glu Gln Gly Val Cys 340 345 350 Ile Arg Lys Asn Trp Asn Ser Ser Asp Tyr Leu His Leu Asn Pro Asp 355 360 365 Asn Phe Ala Ile Gln Leu Glu Lys Gly Gly Lys Phe Thr Val Arg Gly 370 375 380 Lys Pro Thr Leu Glu Asp Leu Glu Gln Phe Ser Glu Lys Phe Tyr Cys 385 390 395 400 Ser Cys Tyr Ser Thr Leu Ser Cys Lys Glu Lys Ala Asp Val Lys Asp 405 410 415 Thr Asp Ala Val Asp Val Cys Ile Ala Asp Gly Val Cys Ile Asp Ala 420 425 430 Phe Leu Lys Pro Pro Met Glu Thr Glu Glu Pro Gln Ile Phe Tyr Asn 435 440 445 Ala Ser Pro Ser Thr Leu Ser 450 455 ＜210＞ 9 ＜211＞ 455 ＜212＞ PRT ＜213＞ Artificial sequence <220> ＜223＞ Synthetic sequence ＜400＞ 9 Leu Asn Phe Arg Ala Pro Pro Val Ile Pro Asn Val Pro Phe Leu Trp 1 5 10 15 Ala Trp Asn Ala Pro Ser Glu Phe Cys Leu Gly Lys Phe Asp Glu Pro 20 25 30 Leu Asp Met Ser Leu Phe Ser Phe Ile Gly Ser Pro Arg Ile Asn Ala 35 40 45 Thr Gly Gln Gly Val Thr Ile Phe Tyr Val Asp Arg Leu Gly Tyr Tyr 50 55 60 Pro Tyr Ile Asp Ser Ile Thr Gly Val Thr Val Asn Gly Gly Ile Pro 65 70 75 80 Gln Lys Ile Ser Leu Gln Asp His Leu Asp Lys Ala Lys Lys Lys Asp Ile 85 90 95 Thr Phe Tyr Met Pro Val Asp Asn Leu Gly Met Ala Val Ile Asp Trp 100 105 110 Glu Glu Trp Arg Pro Thr Trp Ala Arg Asn Trp Lys Pro Lys Asp Val 115 120 125 Tyr Lys Asn Arg Ser Ile Glu Leu Val Gln Gln Gln Asn Val Gln Leu 130 135 140 Ser Leu Thr Glu Ala Thr Glu Lys Ala Lys Gln Glu Phe Glu Lys Ala 145 150 155 160 Gly Lys Asp Phe Leu Val Glu Thr Ile Lys Leu Gly Lys Leu Leu Arg 165 170 175 Pro Asn His Leu Trp Gly Tyr Tyr Leu Phe Pro Asp Cys Tyr Asn His 180 185 190 His Tyr Lys Lys Pro Gly Tyr Asn Gly Ser Cys Phe Asn Val Glu Ile 195 200 205 Lys Arg Asn Asp Asp Leu Ser Trp Leu Trp Asn Glu Ser Thr Ala Leu 210 215 220 Tyr Pro Ser Ile Tyr Leu Asn Thr Gln Gln Ser Pro Val Ala Ala Thr 225 230 235 240 Leu Tyr Val Arg Asn Arg Val Arg Glu Ala Ile Arg Val Ser Lys Ile 245 250 255 Pro Asp Ala Lys Ser Pro Leu Pro Val Phe Ala Tyr Thr Arg Ile Val 260 265 270 Phe Thr Asp Gln Val Leu Lys Phe Leu Ser Gln Asp Glu Leu Val Tyr 275 280 285 Thr Phe Gly Glu Thr Val Ala Leu Gly Ala Ser Gly Ile Val Ile Trp 290 295 300 Gly Thr Leu Ser Ile Thr Arg Thr Lys Glu Ser Cys Gln Ala Ile Lys 305 310 315 320 Glu Tyr Met Asp Thr Thr Leu Asn Pro Tyr Ile Ile Asn Val Thr Leu 325 330 335 Ala Ala Lys Met Cys Ser Gln Val Leu Cys Gln Glu Gln Gly Val Cys 340 345 350 Ile Arg Lys Asn Trp Asn Ser Ser Asp Tyr Leu His Leu Asn Pro Asp 355 360 365 Asn Phe Ala Ile Gln Leu Glu Lys Gly Gly Lys Phe Thr Val Arg Gly 370 375 380 Lys Pro Thr Leu Glu Asp Leu Glu Gln Phe Ser Glu Lys Phe Tyr Cys 385 390 395 400 Ser Cys Tyr Ser Thr Leu Ser Cys Lys Glu Lys Ala Asp Val Lys Asp 405 410 415 Thr Asp Ala Val Asp Val Cys Ile Ala Asp Gly Val Cys Ile Asp Ala 420 425 430 Phe Leu Lys Pro Pro Met Glu Thr Glu Glu Pro Gln Ile Phe Tyr Asn 435 440 445 Ala Ser Pro Ser Thr Leu Ser 450 455 ＜210＞ 10 ＜211＞ 455 ＜212＞ PRT ＜213＞ Artificial sequence <220> ＜223＞ Synthetic sequence ＜400＞ 10 Leu Asn Phe Arg Ala Pro Pro Val Ile Pro Asn Val Pro Phe Leu Trp 1 5 10 15 Ala Trp Asn Ala Pro Ser Glu Phe Cys Leu Gly Lys Phe Asp Glu Pro 20 25 30 Leu Asp Met Ser Leu Phe Ser Phe Ile Gly Ser Pro Arg Ile Asn Ala 35 40 45 Thr Gly Gln Gly Val Thr Ile Phe Tyr Val Asp Arg Leu Gly Tyr Tyr 50 55 60 Pro Tyr Ile Asp Ser Ile Thr Gly Val Thr Val Asn Gly Gly Ile Pro 65 70 75 80 Gln Lys Ile Ser Leu Gln Asp His Leu Asp Lys Ala Lys Lys Lys Asp Ile 85 90 95 Thr Phe Tyr Met Pro Val Asp Asn Leu Gly Met Ala Val Ile Asp Trp 100 105 110 Glu Glu Trp Arg Pro Thr Trp Ala Arg Asn Trp Lys Pro Lys Asp Val 115 120 125 Tyr Lys Asn Arg Ser Ile Glu Leu Val Gln Gln Gln Asn Val Gln Leu 130 135 140 Ser Leu Thr Glu Ala Thr Glu Lys Ala Lys Gln Glu Phe Glu Lys Ala 145 150 155 160 Gly Lys Asp Phe Leu Val Glu Thr Ile Lys Leu Gly Lys Leu Leu Arg 165 170 175 Pro Asn His Leu Trp Gly Tyr Tyr Leu Phe Pro Asp Cys Tyr Asn His 180 185 190 His Tyr Lys Lys Pro Gly Tyr Asn Gly Ser Cys Phe Asn Val Glu Ile 195 200 205 Lys Arg Asn Asp Asp Leu Ser Trp Leu Trp Asn Glu Ser Thr Ala Leu 210 215 220 Tyr Pro Ser Ile Tyr Leu Asn Thr Gln Gln Ser Pro Val Ala Ala Thr 225 230 235 240 Leu Tyr Val Arg Asn Arg Val Arg Glu Ala Ile Arg Val Ser Lys Ile 245 250 255 Pro Asp Ala Lys Ser Pro Leu Pro Val Phe Ala Tyr Thr Arg Ile Val 260 265 270 Phe Thr Asp Gln Val Leu Lys Phe Leu Ser Gln Asp Glu Leu Val Tyr 275 280 285 Thr Phe Gly Glu Thr Val Ala Leu Gly Ala Ser Gly Ile Val Ile Trp 290 295 300 Val Ser Trp Glu Asn Thr Arg Thr Lys Glu Ser Cys Gln Ala Ile Lys 305 310 315 320 Glu Tyr Met Asp Thr Thr Leu Asn Pro Tyr Ile Ile Asn Val Thr Leu 325 330 335 Ala Ala Lys Met Cys Ser Gln Val Leu Cys Gln Glu Gln Gly Val Cys 340 345 350 Ile Arg Lys Asn Trp Asn Ser Ser Asp Tyr Leu His Leu Asn Pro Asp 355 360 365 Asn Phe Ala Ile Gln Leu Glu Lys Gly Gly Lys Phe Thr Val Arg Gly 370 375 380 Lys Pro Thr Leu Glu Asp Leu Glu Gln Phe Ser Glu Lys Phe Tyr Cys 385 390 395 400 Ser Cys Tyr Ser Thr Leu Ser Cys Lys Glu Lys Ala Asp Val Lys Asp 405 410 415 Thr Asp Ala Val Asp Val Cys Ile Ala Asp Gly Val Cys Ile Asp Ala 420 425 430 Phe Leu Lys Pro Pro Met Glu Thr Glu Glu Pro Gln Ile Phe Tyr Asn 435 440 445 Ala Ser Pro Ser Thr Leu Ser 450 455 ＜210＞ 11 ＜211＞ 455 ＜212＞ PRT ＜213＞ Artificial sequence <220> ＜223＞ Synthetic sequence ＜400＞ 11 Leu Asn Phe Arg Ala Pro Pro Val Ile Pro Asn Val Pro Phe Leu Trp 1 5 10 15 Ala Trp Asn Ala Pro Ser Glu Phe Cys Leu Gly Lys Phe Asp Glu Pro 20 25 30 Leu Asp Met Ser Leu Phe Ser Phe Ile Gly Ser Pro Arg Ile Asn Ala 35 40 45 Thr Gly Gln Gly Val Thr Ile Phe Tyr Val Asp Arg Leu Gly Tyr Tyr 50 55 60 Pro Tyr Ile Asp Ser Ile Thr Gly Val Thr Val Asn Gly Gly Ile Pro 65 70 75 80 Gln Lys Ile Ser Leu Gln Asp His Leu Asp Lys Ala Lys Lys Lys Asp Ile 85 90 95 Thr Phe Tyr Met Pro Val Asp Asn Leu Gly Met Ala Val Ile Asp Trp 100 105 110 Glu Glu Trp Arg Pro Thr Trp Ala Arg Asn Trp Lys Pro Lys Asp Val 115 120 125 Tyr Lys Asn Arg Ser Ile Glu Leu Val Gln Gln Gln Asn Val Gln Leu 130 135 140 Ser Leu Thr Glu Ala Thr Glu Lys Ala Lys Gln Glu Phe Glu Lys Ala 145 150 155 160 Gly Lys Asp Phe Leu Val Glu Thr Ile Lys Leu Gly Lys Leu Leu Arg 165 170 175 Pro Asn His Leu Trp Gly Tyr Tyr Leu Phe Pro Asp Cys Tyr Asn His 180 185 190 His Tyr Lys Lys Pro Gly Tyr Asn Gly Ser Cys Phe Asn Val Glu Ile 195 200 205 Lys Arg Asn Asp Asp Leu Ser Trp Leu Trp Asn Glu Ser Thr Ala Leu 210 215 220 Tyr Pro Ser Ile Tyr Leu Asn Thr Gln Gln Ser Pro Val Ala Ala Thr 225 230 235 240 Leu Tyr Val Arg Asn Arg Val Arg Glu Ala Ile Arg Val Ser Lys Ile 245 250 255 Pro Asp Ala Lys Ser Pro Leu Pro Val Phe Ala Tyr Thr Arg Ile Val 260 265 270 Phe Thr Asp Gln Val Leu Lys Phe Leu Ser Gln Asp Glu Leu Val Tyr 275 280 285 Thr Phe Gly Glu Thr Val Ala Leu Gly Ala Ser Gly Ile Val Ile Trp 290 295 300 Gly Thr Leu Ser Asn Thr Arg Thr Lys Glu Ser Cys Gln Ala Ile Lys 305 310 315 320 Glu Tyr Met Asp Thr Thr Leu Asn Pro Tyr Ile Ile Asn Val Thr Leu 325 330 335 Ala Ala Lys Met Cys Ser Gln Val Leu Cys Gln Glu Gln Gly Val Cys 340 345 350 Ile Arg Lys Asn Trp Asn Ser Ser Asp Tyr Leu His Leu Asn Pro Asp 355 360 365 Asn Phe Ala Ile Gln Leu Glu Lys Gly Gly Lys Phe Thr Val Arg Gly 370 375 380 Lys Pro Thr Leu Glu Asp Leu Glu Gln Phe Ser Glu Lys Phe Tyr Cys 385 390 395 400 Ser Cys Tyr Ser Thr Leu Ser Cys Lys Glu Lys Ala Asp Val Lys Asp 405 410 415 Thr Asp Ala Val Asp Val Cys Ile Ala Asp Gly Val Cys Ile Asp Ala 420 425 430 Phe Leu Lys Pro Pro Met Glu Thr Glu Glu Pro Gln Ile Phe Tyr Asn 435 440 445 Ala Ser Pro Ser Thr Leu Ser 450 455 ＜210＞ 12 ＜211＞ 455 ＜212＞ PRT ＜213＞ Artificial sequence <220> ＜223＞ Synthetic sequence ＜400＞ 12 Leu Asn Phe Arg Ala Pro Pro Val Ile Pro Asn Val Pro Phe Leu Trp 1 5 10 15 Ala Trp Asn Ala Pro Ser Glu Phe Cys Leu Gly Lys Phe Asp Glu Pro 20 25 30 Leu Asp Met Ser Leu Phe Ser Phe Ile Gly Ser Pro Arg Ile Asn Ala 35 40 45 Thr Gly Gln Gly Val Thr Ile Phe Tyr Val Asp Arg Leu Gly Tyr Tyr 50 55 60 Pro Tyr Ile Asp Ser Ile Thr Gly Val Thr Val Asn Gly Gly Ile Pro 65 70 75 80 Gln Lys Ile Ser Leu Gln Asp His Leu Asp Lys Ala Lys Lys Lys Asp Ile 85 90 95 Thr Phe Tyr Met Pro Val Asp Asn Leu Gly Met Ala Val Ile Asp Trp 100 105 110 Glu Glu Trp Arg Pro Thr Trp Ala Arg Asn Trp Lys Pro Lys Asp Val 115 120 125 Tyr Lys Asn Arg Ser Ile Glu Leu Val Gln Gln Gln Asn Val Gln Leu 130 135 140 Ser Leu Thr Glu Ala Thr Glu Lys Ala Lys Gln Glu Phe Glu Lys Ala 145 150 155 160 Gly Lys Asp Phe Leu Val Glu Thr Ile Lys Leu Gly Lys Leu Leu Arg 165 170 175 Pro Asn His Leu Trp Gly Tyr Tyr Leu Phe Pro Asp Cys Tyr Asn His 180 185 190 His Tyr Lys Lys Pro Gly Tyr Asn Gly Ser Cys Phe Asn Val Glu Ile 195 200 205 Lys Arg Asn Asp Asp Leu Ser Trp Leu Trp Asn Glu Ser Thr Ala Leu 210 215 220 Tyr Pro Ser Ile Tyr Leu Asn Thr Gln Gln Ser Pro Val Ala Ala Thr 225 230 235 240 Leu Tyr Val Arg Asn Arg Val Arg Glu Ala Ile Arg Val Ser Lys Ile 245 250 255 Pro Asp Ala Lys Ser Pro Leu Pro Val Phe Ala Tyr Thr Arg Ile Val 260 265 270 Phe Thr Asp Gln Val Leu Lys Phe Leu Ser Gln Asp Glu Leu Val Tyr 275 280 285 Thr Phe Gly Glu Thr Val Ala Leu Gly Ala Ser Gly Ile Val Ile Trp 290 295 300 Gly Thr Leu Glu Asn Thr Arg Thr Lys Glu Ser Cys Gln Ala Ile Lys 305 310 315 320 Glu Tyr Met Asp Thr Thr Leu Asn Pro Tyr Ile Ile Asn Val Thr Leu 325 330 335 Ala Ala Lys Met Cys Ser Gln Val Leu Cys Gln Glu Gln Gly Val Cys 340 345 350 Ile Arg Lys Asn Trp Asn Ser Ser Asp Tyr Leu His Leu Asn Pro Asp 355 360 365 Asn Phe Ala Ile Gln Leu Glu Lys Gly Gly Lys Phe Thr Val Arg Gly 370 375 380 Lys Pro Thr Leu Glu Asp Leu Glu Gln Phe Ser Glu Lys Phe Tyr Cys 385 390 395 400 Ser Cys Tyr Ser Thr Leu Ser Cys Lys Glu Lys Ala Asp Val Lys Asp 405 410 415 Thr Asp Ala Val Asp Val Cys Ile Ala Asp Gly Val Cys Ile Asp Ala 420 425 430 Phe Leu Lys Pro Pro Met Glu Thr Glu Glu Pro Gln Ile Phe Tyr Asn 435 440 445 Ala Ser Pro Ser Thr Leu Ser 450 455 ＜210＞ 13 ＜211＞ 455 ＜212＞ PRT ＜213＞ Artificial sequence <220> ＜223＞ Synthetic sequence ＜400＞ 13 Leu Asn Phe Arg Ala Pro Pro Val Ile Pro Asn Val Pro Phe Leu Trp 1 5 10 15 Ala Trp Asn Ala Pro Ser Glu Phe Cys Leu Gly Lys Phe Asp Glu Pro 20 25 30 Leu Asp Met Ser Leu Phe Ser Phe Ile Gly Ser Pro Arg Ile Asn Ala 35 40 45 Thr Gly Gln Gly Val Thr Ile Phe Tyr Val Asp Arg Leu Gly Tyr Tyr 50 55 60 Pro Tyr Ile Asp Ser Ile Thr Gly Val Thr Val Asn Gly Gly Ile Pro 65 70 75 80 Gln Lys Ile Ser Leu Gln Asp His Leu Asp Lys Ala Lys Lys Lys Asp Ile 85 90 95 Thr Phe Tyr Met Pro Val Asp Asn Leu Gly Met Ala Val Ile Asp Trp 100 105 110 Glu Glu Trp Arg Pro Thr Trp Ala Arg Asn Trp Lys Pro Lys Asp Val 115 120 125 Tyr Lys Asn Arg Ser Ile Glu Leu Val Gln Gln Gln Asn Val Gln Leu 130 135 140 Ser Leu Thr Glu Ala Thr Glu Lys Ala Lys Gln Glu Phe Glu Lys Ala 145 150 155 160 Gly Lys Asp Phe Leu Val Glu Thr Ile Lys Leu Gly Lys Leu Leu Arg 165 170 175 Pro Asn His Leu Trp Gly Tyr Tyr Leu Phe Pro Asp Cys Tyr Asn His 180 185 190 His Tyr Lys Lys Pro Gly Tyr Asn Gly Ser Cys Phe Asn Val Glu Ile 195 200 205 Lys Arg Asn Asp Asp Leu Ser Trp Leu Trp Asn Glu Ser Thr Ala Leu 210 215 220 Tyr Pro Ser Ile Tyr Leu Asn Thr Gln Gln Ser Pro Val Ala Ala Thr 225 230 235 240 Leu Tyr Val Arg Asn Arg Val Arg Glu Ala Ile Arg Val Ser Lys Ile 245 250 255 Pro Asp Ala Lys Ser Pro Leu Pro Val Phe Ala Tyr Thr Arg Ile Val 260 265 270 Phe Thr Asp Gln Val Leu Lys Phe Leu Ser Gln Asp Glu Leu Val Tyr 275 280 285 Thr Phe Gly Glu Thr Val Ala Leu Gly Ala Ser Gly Ile Val Ile Trp 290 295 300 Gly Thr Trp Glu Asn Thr Arg Thr Lys Glu Ser Cys Gln Ala Ile Lys 305 310 315 320 Glu Tyr Met Asp Thr Thr Leu Asn Pro Tyr Ile Ile Asn Val Thr Leu 325 330 335 Ala Ala Lys Met Cys Ser Gln Val Leu Cys Gln Glu Gln Gly Val Cys 340 345 350 Ile Arg Lys Asn Trp Asn Ser Ser Asp Tyr Leu His Leu Asn Pro Asp 355 360 365 Asn Phe Ala Ile Gln Leu Glu Lys Gly Gly Lys Phe Thr Val Arg Gly 370 375 380 Lys Pro Thr Leu Glu Asp Leu Glu Gln Phe Ser Glu Lys Phe Tyr Cys 385 390 395 400 Ser Cys Tyr Ser Thr Leu Ser Cys Lys Glu Lys Ala Asp Val Lys Asp 405 410 415 Thr Asp Ala Val Asp Val Cys Ile Ala Asp Gly Val Cys Ile Asp Ala 420 425 430 Phe Leu Lys Pro Pro Met Glu Thr Glu Glu Pro Gln Ile Phe Tyr Asn 435 440 445 Ala Ser Pro Ser Thr Leu Ser 450 455 ＜210＞ 14 ＜211＞ 455 ＜212＞ PRT ＜213＞ Artificial sequence <220> ＜223＞ Synthetic sequence ＜400＞ 14 Leu Asn Phe Arg Ala Pro Pro Val Ile Pro Asn Val Pro Phe Leu Trp 1 5 10 15 Ala Trp Asn Ala Pro Ser Glu Phe Cys Leu Gly Lys Phe Asp Glu Pro 20 25 30 Leu Asp Met Ser Leu Phe Ser Phe Ile Gly Ser Pro Arg Ile Asn Ala 35 40 45 Thr Gly Gln Gly Val Thr Ile Phe Tyr Val Asp Arg Leu Gly Tyr Tyr 50 55 60 Pro Tyr Ile Asp Ser Ile Thr Gly Val Thr Val Asn Gly Gly Ile Pro 65 70 75 80 Gln Lys Ile Ser Leu Gln Asp His Leu Asp Lys Ala Lys Lys Lys Asp Ile 85 90 95 Thr Phe Tyr Met Pro Val Asp Asn Leu Gly Met Ala Val Ile Asp Trp 100 105 110 Glu Glu Trp Arg Pro Thr Trp Ala Arg Asn Trp Lys Pro Lys Asp Val 115 120 125 Tyr Lys Asn Arg Ser Ile Glu Leu Val Gln Gln Gln Asn Val Gln Leu 130 135 140 Ser Leu Thr Glu Ala Thr Glu Lys Ala Lys Gln Glu Phe Glu Lys Ala 145 150 155 160 Gly Lys Asp Phe Leu Val Glu Thr Ile Lys Leu Gly Lys Leu Leu Arg 165 170 175 Pro Asn His Leu Trp Gly Tyr Tyr Leu Phe Pro Asp Cys Tyr Asn His 180 185 190 His Tyr Lys Lys Pro Gly Tyr Asn Gly Ser Cys Phe Asn Val Glu Ile 195 200 205 Lys Arg Asn Asp Asp Leu Ser Trp Leu Trp Asn Glu Ser Thr Ala Leu 210 215 220 Tyr Pro Ser Ile Tyr Leu Asn Thr Gln Gln Ser Pro Val Ala Ala Thr 225 230 235 240 Leu Tyr Val Arg Asn Arg Val Arg Glu Ala Ile Arg Val Ser Lys Ile 245 250 255 Pro Asp Ala Lys Ser Pro Leu Pro Val Phe Ala Tyr Thr Arg Ile Val 260 265 270 Phe Thr Asp Gln Val Leu Lys Phe Leu Ser Gln Asp Glu Leu Val Tyr 275 280 285 Thr Phe Gly Glu Thr Val Ala Leu Gly Ala Ser Gly Ile Val Ile Trp 290 295 300 Gly Thr Leu Ser Ile Thr Arg Thr Lys Glu Ser Cys Gln Ala Ile Lys 305 310 315 320 Glu Tyr Met Asp Thr Thr Leu Asn Pro Phe Ile Leu Asn Val Thr Leu 325 330 335 Ala Ala Lys Met Cys Ser Gln Val Leu Cys Gln Glu Gln Gly Val Cys 340 345 350 Ile Arg Lys Asn Trp Asn Ser Ser Asp Tyr Leu His Leu Asn Pro Asp 355 360 365 Asn Phe Ala Ile Gln Leu Glu Lys Gly Gly Lys Phe Thr Val Arg Gly 370 375 380 Lys Pro Thr Leu Glu Asp Leu Glu Gln Phe Ser Glu Lys Phe Tyr Cys 385 390 395 400 Ser Cys Tyr Ser Thr Leu Ser Cys Lys Glu Lys Ala Asp Val Lys Asp 405 410 415 Thr Asp Ala Val Asp Val Cys Ile Ala Asp Gly Val Cys Ile Asp Ala 420 425 430 Phe Leu Lys Pro Pro Met Glu Thr Glu Glu Pro Gln Ile Phe Tyr Asn 435 440 445 Ala Ser Pro Ser Thr Leu Ser 450 455 ＜210＞ 15 ＜211＞ 455 ＜212＞ PRT ＜213＞ Artificial sequence <220> ＜223＞ Synthetic sequence ＜400＞ 15 Leu Asn Phe Arg Ala Pro Pro Val Ile Pro Asn Val Pro Phe Leu Trp 1 5 10 15 Ala Trp Asn Ala Pro Ser Glu Phe Cys Leu Gly Lys Phe Asp Glu Pro 20 25 30 Leu Asp Met Ser Leu Phe Ser Phe Ile Gly Ser Pro Arg Ile Asn Ala 35 40 45 Thr Gly Gln Gly Val Thr Ile Phe Tyr Val Asp Arg Leu Gly Tyr Tyr 50 55 60 Pro Tyr Ile Asp Ser Ile Thr Gly Val Thr Val Asn Gly Gly Ile Pro 65 70 75 80 Gln Lys Ile Ser Leu Gln Asp His Leu Asp Lys Ala Lys Lys Lys Asp Ile 85 90 95 Thr Phe Tyr Met Pro Val Asp Asn Leu Gly Met Ala Val Ile Asp Trp 100 105 110 Glu Glu Trp Arg Pro Thr Trp Ala Arg Asn Trp Lys Pro Lys Asp Val 115 120 125 Tyr Lys Asn Arg Ser Ile Glu Leu Val Gln Gln Gln Asn Val Gln Leu 130 135 140 Ser Leu Thr Glu Ala Thr Glu Lys Ala Lys Gln Glu Phe Glu Lys Ala 145 150 155 160 Gly Lys Asp Phe Leu Val Glu Thr Ile Lys Leu Gly Lys Leu Leu Arg 165 170 175 Pro Asn His Leu Trp Gly Tyr Tyr Leu Phe Pro Asp Cys Tyr Asn His 180 185 190 His Tyr Lys Lys Pro Gly Tyr Asn Gly Ser Cys Phe Asn Val Glu Ile 195 200 205 Lys Arg Asn Asp Asp Leu Ser Trp Leu Trp Asn Glu Ser Thr Ala Leu 210 215 220 Tyr Pro Ser Ile Tyr Leu Asn Thr Gln Gln Ser Pro Val Ala Ala Thr 225 230 235 240 Leu Tyr Val Arg Asn Arg Val Arg Glu Ala Ile Arg Val Ser Lys Ile 245 250 255 Pro Asp Ala Lys Ser Pro Leu Pro Val Phe Ala Tyr Thr Arg Ile Val 260 265 270 Phe Thr Asp Gln Val Leu Lys Phe Leu Ser Gln Asp Glu Leu Val Tyr 275 280 285 Thr Phe Gly Glu Thr Val Ala Leu Gly Ala Ser Gly Ile Val Ile Trp 290 295 300 Gly Thr Leu Ser Ile Thr Arg Thr Lys Glu Ser Cys Gln Ala Ile Lys 305 310 315 320 Glu Tyr Met Asp Thr Thr Leu Asn Pro Phe Ile Leu Asn Val Thr Ser 325 330 335 Gly Ala Lys Met Cys Ser Gln Val Leu Cys Gln Glu Gln Gly Val Cys 340 345 350 Ile Arg Lys Asn Trp Asn Ser Ser Asp Tyr Leu His Leu Asn Pro Asp 355 360 365 Asn Phe Ala Ile Gln Leu Glu Lys Gly Gly Lys Phe Thr Val Arg Gly 370 375 380 Lys Pro Thr Leu Glu Asp Leu Glu Gln Phe Ser Glu Lys Phe Tyr Cys 385 390 395 400 Ser Cys Tyr Ser Thr Leu Ser Cys Lys Glu Lys Ala Asp Val Lys Asp 405 410 415 Thr Asp Ala Val Asp Val Cys Ile Ala Asp Gly Val Cys Ile Asp Ala 420 425 430 Phe Leu Lys Pro Pro Met Glu Thr Glu Glu Pro Gln Ile Phe Tyr Asn 435 440 445 Ala Ser Pro Ser Thr Leu Ser 450 455 ＜210＞ 16 <211> 453 ＜212＞ PRT ＜213＞ Artificial sequence <220> ＜223＞ Synthetic sequence ＜400＞ 16 Phe Arg Ala Pro Pro Val Ile Pro Asn Val Pro Phe Leu Trp Ala Trp 1 5 10 15 Asn Ala Pro Ser Glu Phe Cys Leu Gly Lys Phe Asp Glu Pro Leu Asp 20 25 30 Met Ser Leu Phe Ser Phe Ile Gly Ser Pro Arg Ile Asn Ala Thr Gly 35 40 45 Gln Gly Val Thr Ile Phe Tyr Val Asp Arg Leu Gly Tyr Tyr Pro Tyr 50 55 60 Ile Asp Ser Ile Thr Gly Val Thr Val Asn Gly Gly Ile Pro Gln Lys 65 70 75 80 Ile Ser Leu Gln Asp His Leu Asp Lys Ala Lys Lys Asp Ile Thr Phe 85 90 95 Tyr Met Pro Val Asp Asn Leu Gly Met Ala Val Ile Asp Trp Glu Glu 100 105 110 Trp Arg Pro Thr Trp Ala Arg Asn Trp Lys Pro Lys Asp Val Tyr Lys 115 120 125 Asn Arg Ser Ile Glu Leu Val Gln Gln Gln Asn Val Gln Leu Ser Leu 130 135 140 Thr Glu Ala Thr Glu Lys Ala Lys Gln Glu Phe Glu Lys Ala Gly Lys 145 150 155 160 Asp Phe Leu Val Glu Thr Ile Lys Leu Gly Lys Leu Leu Arg Pro Asn 165 170 175 His Leu Trp Gly Tyr Tyr Leu Phe Pro Asp Cys Tyr Asn His His Tyr 180 185 190 Lys Lys Pro Gly Tyr Asn Gly Ser Cys Phe Asn Val Glu Ile Lys Arg 195 200 205 Asn Asp Asp Leu Ser Trp Leu Trp Asn Glu Ser Thr Ala Leu Tyr Pro 210 215 220 Ser Ile Tyr Leu Asn Thr Gln Gln Ser Pro Val Ala Ala Thr Leu Tyr 225 230 235 240 Val Arg Asn Arg Val Arg Glu Ala Ile Arg Val Ser Lys Ile Pro Asp 245 250 255 Ala Lys Ser Pro Leu Pro Val Phe Ala Tyr Thr Arg Ile Val Phe Thr 260 265 270 Asp Gln Val Leu Lys Phe Leu Ser Gln Asp Glu Leu Val Tyr Thr Phe 275 280 285 Gly Glu Thr Val Ala Leu Gly Ala Ser Gly Ile Val Ile Trp Gly Thr 290 295 300 Leu Ser Ile Thr Arg Thr Lys Glu Ser Cys Gln Ala Ile Lys Glu Tyr 305 310 315 320 Met Asp Thr Thr Leu Asn Pro Tyr Ile Ile Asn Val Thr Leu Ala Ala 325 330 335 Lys Met Cys Ser Gln Val Leu Cys Gln Glu Gln Gly Val Cys Ile Arg 340 345 350 Lys Asn Trp Asn Ser Ser Asp Tyr Leu His Leu Asn Pro Asp Asn Phe 355 360 365 Ala Ile Gln Leu Glu Lys Gly Gly Lys Phe Thr Val Arg Gly Lys Pro 370 375 380 Thr Leu Glu Asp Leu Glu Gln Phe Ser Glu Lys Phe Tyr Cys Ser Cys 385 390 395 400 Tyr Ser Thr Leu Ser Cys Lys Glu Lys Ala Asp Val Lys Asp Thr Asp 405 410 415 Ala Val Asp Val Cys Ile Ala Asp Gly Val Cys Ile Asp Ala Phe Leu 420 425 430 Lys Pro Pro Met Glu Thr Glu Glu Pro Gln Ile Phe Tyr Asn Ala Ser 435 440 445 Pro Ser Thr Leu Ser 450 ＜210＞ 17 ＜211＞ 430 ＜212＞ PRT ＜213＞ Artificial sequence <220> ＜223＞ Synthetic sequence ＜400＞ 17 Leu Asn Phe Arg Ala Pro Pro Val Ile Pro Asn Val Pro Phe Leu Trp 1 5 10 15 Ala Trp Asn Ala Pro Ser Glu Phe Cys Leu Gly Lys Phe Asp Glu Pro 20 25 30 Leu Asp Met Ser Leu Phe Ser Phe Ile Gly Ser Pro Arg Ile Asn Ala 35 40 45 Thr Gly Gln Gly Val Thr Ile Phe Tyr Val Asp Arg Leu Gly Tyr Tyr 50 55 60 Pro Tyr Ile Asp Ser Ile Thr Gly Val Thr Val Asn Gly Gly Ile Pro 65 70 75 80 Gln Lys Ile Ser Leu Gln Asp His Leu Asp Lys Ala Lys Lys Lys Asp Ile 85 90 95 Thr Phe Tyr Met Pro Val Asp Asn Leu Gly Met Ala Val Ile Asp Trp 100 105 110 Glu Glu Trp Arg Pro Thr Trp Ala Arg Asn Trp Lys Pro Lys Asp Val 115 120 125 Tyr Lys Asn Arg Ser Ile Glu Leu Val Gln Gln Gln Asn Val Gln Leu 130 135 140 Ser Leu Thr Glu Ala Thr Glu Lys Ala Lys Gln Glu Phe Glu Lys Ala 145 150 155 160 Gly Lys Asp Phe Leu Val Glu Thr Ile Lys Leu Gly Lys Leu Leu Arg 165 170 175 Pro Asn His Leu Trp Gly Tyr Tyr Leu Phe Pro Asp Cys Tyr Asn His 180 185 190 His Tyr Lys Lys Pro Gly Tyr Asn Gly Ser Cys Phe Asn Val Glu Ile 195 200 205 Lys Arg Asn Asp Asp Leu Ser Trp Leu Trp Asn Glu Ser Thr Ala Leu 210 215 220 Tyr Pro Ser Ile Tyr Leu Asn Thr Gln Gln Ser Pro Val Ala Ala Thr 225 230 235 240 Leu Tyr Val Arg Asn Arg Val Arg Glu Ala Ile Arg Val Ser Lys Ile 245 250 255 Pro Asp Ala Lys Ser Pro Leu Pro Val Phe Ala Tyr Thr Arg Ile Val 260 265 270 Phe Thr Asp Gln Val Leu Lys Phe Leu Ser Gln Asp Glu Leu Val Tyr 275 280 285 Thr Phe Gly Glu Thr Val Ala Leu Gly Ala Ser Gly Ile Val Ile Trp 290 295 300 Gly Thr Leu Ser Ile Thr Arg Thr Lys Glu Ser Cys Gln Ala Ile Lys 305 310 315 320 Glu Tyr Met Asp Thr Thr Leu Asn Pro Tyr Ile Ile Asn Val Thr Leu 325 330 335 Ala Ala Lys Met Cys Ser Gln Val Leu Cys Gln Glu Gln Gly Val Cys 340 345 350 Ile Arg Lys Asn Trp Asn Ser Ser Asp Tyr Leu His Leu Asn Pro Asp 355 360 365 Asn Phe Ala Ile Gln Leu Glu Lys Gly Gly Lys Phe Thr Val Arg Gly 370 375 380 Lys Pro Thr Leu Glu Asp Leu Glu Gln Phe Ser Glu Lys Phe Tyr Cys 385 390 395 400 Ser Cys Tyr Ser Thr Leu Ser Cys Lys Glu Lys Ala Asp Val Lys Asp 405 410 415 Thr Asp Ala Val Asp Val Cys Ile Ala Asp Gly Val Cys Ile 420 425 430 ＜210＞ 18 <211> 433 ＜212＞ PRT ＜213＞ Artificial sequence <220> ＜223＞ Synthetic sequence ＜400＞ 18 Leu Asn Phe Arg Ala Pro Pro Val Ile Pro Asn Val Pro Phe Leu Trp 1 5 10 15 Ala Trp Asn Ala Pro Ser Glu Phe Cys Leu Gly Lys Phe Asp Glu Pro 20 25 30 Leu Asp Met Ser Leu Phe Ser Phe Ile Gly Ser Pro Arg Ile Asn Ala 35 40 45 Thr Gly Gln Gly Val Thr Ile Phe Tyr Val Asp Arg Leu Gly Tyr Tyr 50 55 60 Pro Tyr Ile Asp Ser Ile Thr Gly Val Thr Val Asn Gly Gly Ile Pro 65 70 75 80 Gln Lys Ile Ser Leu Gln Asp His Leu Asp Lys Ala Lys Lys Lys Asp Ile 85 90 95 Thr Phe Tyr Met Pro Val Asp Asn Leu Gly Met Ala Val Ile Asp Trp 100 105 110 Glu Glu Trp Arg Pro Thr Trp Ala Arg Asn Trp Lys Pro Lys Asp Val 115 120 125 Tyr Lys Asn Arg Ser Ile Glu Leu Val Gln Gln Gln Asn Val Gln Leu 130 135 140 Ser Leu Thr Glu Ala Thr Glu Lys Ala Lys Gln Glu Phe Glu Lys Ala 145 150 155 160 Gly Lys Asp Phe Leu Val Glu Thr Ile Lys Leu Gly Lys Leu Leu Arg 165 170 175 Pro Asn His Leu Trp Gly Tyr Tyr Leu Phe Pro Asp Cys Tyr Asn His 180 185 190 His Tyr Lys Lys Pro Gly Tyr Asn Gly Ser Cys Phe Asn Val Glu Ile 195 200 205 Lys Arg Asn Asp Asp Leu Ser Trp Leu Trp Asn Glu Ser Thr Ala Leu 210 215 220 Tyr Pro Ser Ile Tyr Leu Asn Thr Gln Gln Ser Pro Val Ala Ala Thr 225 230 235 240 Leu Tyr Val Arg Asn Arg Val Arg Glu Ala Ile Arg Val Ser Lys Ile 245 250 255 Pro Asp Ala Lys Ser Pro Leu Pro Val Phe Ala Tyr Thr Arg Ile Val 260 265 270 Phe Thr Asp Gln Val Leu Lys Phe Leu Ser Gln Asp Glu Leu Val Tyr 275 280 285 Thr Phe Gly Glu Thr Val Ala Leu Gly Ala Ser Gly Ile Val Ile Trp 290 295 300 Gly Thr Leu Ser Ile Thr Arg Thr Lys Glu Ser Cys Gln Ala Ile Lys 305 310 315 320 Glu Tyr Met Asp Thr Thr Leu Asn Pro Tyr Ile Ile Asn Val Thr Leu 325 330 335 Ala Ala Lys Met Cys Ser Gln Val Leu Cys Gln Glu Gln Gly Val Cys 340 345 350 Ile Arg Lys Asn Trp Asn Ser Ser Asp Tyr Leu His Leu Asn Pro Asp 355 360 365 Asn Phe Ala Ile Gln Leu Glu Lys Gly Gly Lys Phe Thr Val Arg Gly 370 375 380 Lys Pro Thr Leu Glu Asp Leu Glu Gln Phe Ser Glu Lys Phe Tyr Cys 385 390 395 400 Ser Cys Tyr Ser Thr Leu Ser Cys Lys Glu Lys Ala Asp Val Lys Asp 405 410 415 Thr Asp Ala Val Asp Val Cys Ile Ala Asp Gly Val Cys Ile Asp Ala 420 425 430 Phe ＜210＞ 19 ＜211＞ 436 ＜212＞ PRT ＜213＞ Artificial sequence <220> ＜223＞ Synthetic sequence ＜400＞ 19 Leu Asn Phe Arg Ala Pro Pro Val Ile Pro Asn Val Pro Phe Leu Trp 1 5 10 15 Ala Trp Asn Ala Pro Ser Glu Phe Cys Leu Gly Lys Phe Asp Glu Pro 20 25 30 Leu Asp Met Ser Leu Phe Ser Phe Ile Gly Ser Pro Arg Ile Asn Ala 35 40 45 Thr Gly Gln Gly Val Thr Ile Phe Tyr Val Asp Arg Leu Gly Tyr Tyr 50 55 60 Pro Tyr Ile Asp Ser Ile Thr Gly Val Thr Val Asn Gly Gly Ile Pro 65 70 75 80 Gln Lys Ile Ser Leu Gln Asp His Leu Asp Lys Ala Lys Lys Lys Asp Ile 85 90 95 Thr Phe Tyr Met Pro Val Asp Asn Leu Gly Met Ala Val Ile Asp Trp 100 105 110 Glu Glu Trp Arg Pro Thr Trp Ala Arg Asn Trp Lys Pro Lys Asp Val 115 120 125 Tyr Lys Asn Arg Ser Ile Glu Leu Val Gln Gln Gln Asn Val Gln Leu 130 135 140 Ser Leu Thr Glu Ala Thr Glu Lys Ala Lys Gln Glu Phe Glu Lys Ala 145 150 155 160 Gly Lys Asp Phe Leu Val Glu Thr Ile Lys Leu Gly Lys Leu Leu Arg 165 170 175 Pro Asn His Leu Trp Gly Tyr Tyr Leu Phe Pro Asp Cys Tyr Asn His 180 185 190 His Tyr Lys Lys Pro Gly Tyr Asn Gly Ser Cys Phe Asn Val Glu Ile 195 200 205 Lys Arg Asn Asp Asp Leu Ser Trp Leu Trp Asn Glu Ser Thr Ala Leu 210 215 220 Tyr Pro Ser Ile Tyr Leu Asn Thr Gln Gln Ser Pro Val Ala Ala Thr 225 230 235 240 Leu Tyr Val Arg Asn Arg Val Arg Glu Ala Ile Arg Val Ser Lys Ile 245 250 255 Pro Asp Ala Lys Ser Pro Leu Pro Val Phe Ala Tyr Thr Arg Ile Val 260 265 270 Phe Thr Asp Gln Val Leu Lys Phe Leu Ser Gln Asp Glu Leu Val Tyr 275 280 285 Thr Phe Gly Glu Thr Val Ala Leu Gly Ala Ser Gly Ile Val Ile Trp 290 295 300 Gly Thr Leu Ser Ile Thr Arg Thr Lys Glu Ser Cys Gln Ala Ile Lys 305 310 315 320 Glu Tyr Met Asp Thr Thr Leu Asn Pro Tyr Ile Ile Asn Val Thr Leu 325 330 335 Ala Ala Lys Met Cys Ser Gln Val Leu Cys Gln Glu Gln Gly Val Cys 340 345 350 Ile Arg Lys Asn Trp Asn Ser Ser Asp Tyr Leu His Leu Asn Pro Asp 355 360 365 Asn Phe Ala Ile Gln Leu Glu Lys Gly Gly Lys Phe Thr Val Arg Gly 370 375 380 Lys Pro Thr Leu Glu Asp Leu Glu Gln Phe Ser Glu Lys Phe Tyr Cys 385 390 395 400 Ser Cys Tyr Ser Thr Leu Ser Cys Lys Glu Lys Ala Asp Val Lys Asp 405 410 415 Thr Asp Ala Val Asp Val Cys Ile Ala Asp Gly Val Cys Ile Asp Ala 420 425 430 Phe Leu Lys Pro 435 ＜210＞ 20 ＜211＞ 452 ＜212＞ PRT ＜213＞ Artificial sequence <220> ＜223＞ Synthetic sequence ＜400＞ 20 Phe Arg Gly Pro Leu Leu Pro Asn Arg Pro Phe Leu Trp Ala Trp Asn 1 5 10 15 Ala Pro Ser Glu Phe Cys Leu Gly Lys Phe Asp Glu Pro Leu Asp Met 20 25 30 Ser Leu Phe Ser Phe Ile Gly Ser Pro Arg Ile Asn Ala Thr Gly Gln 35 40 45 Gly Val Thr Ile Phe Tyr Val Asp Arg Leu Gly Tyr Tyr Pro Tyr Ile 50 55 60 Asp Ser Ile Thr Gly Val Thr Val Asn Gly Gly Ile Pro Gln Lys Ile 65 70 75 80 Ser Leu Gln Asp His Leu Asp Lys Ala Lys Lys Asp Ile Thr Phe Tyr 85 90 95 Met Pro Val Asp Asn Leu Gly Met Ala Val Ile Asp Trp Glu Glu Trp 100 105 110 Arg Pro Thr Trp Ala Arg Asn Trp Lys Pro Lys Asp Val Tyr Lys Asn 115 120 125 Arg Ser Ile Glu Leu Val Gln Gln Gln Asn Val Gln Leu Ser Leu Thr 130 135 140 Glu Ala Thr Glu Lys Ala Lys Gln Glu Phe Glu Lys Ala Gly Lys Asp 145 150 155 160 Phe Leu Val Glu Thr Ile Lys Leu Gly Lys Leu Leu Arg Pro Asn His 165 170 175 Leu Trp Gly Tyr Tyr Leu Phe Pro Asp Cys Tyr Asn His His Tyr Lys 180 185 190 Lys Pro Gly Tyr Asn Gly Ser Cys Phe Asn Val Glu Ile Lys Arg Asn 195 200 205 Asp Asp Leu Ser Trp Leu Trp Asn Glu Ser Thr Ala Leu Tyr Pro Ser 210 215 220 Ile Tyr Leu Asn Thr Gln Gln Ser Pro Val Ala Ala Thr Leu Tyr Val 225 230 235 240 Arg Asn Arg Val Arg Glu Ala Ile Arg Val Ser Lys Ile Pro Asp Ala 245 250 255 Lys Ser Pro Leu Pro Val Phe Ala Tyr Thr Arg Ile Val Phe Thr Asp 260 265 270 Gln Val Leu Lys Phe Leu Ser Gln Asp Glu Leu Val Tyr Thr Phe Gly 275 280 285 Glu Thr Val Ala Leu Gly Ala Ser Gly Ile Val Ile Trp Gly Thr Leu 290 295 300 Ser Ile Thr Arg Thr Lys Glu Ser Cys Gln Ala Ile Lys Glu Tyr Met 305 310 315 320 Asp Thr Thr Leu Asn Pro Tyr Ile Ile Asn Val Thr Leu Ala Ala Lys 325 330 335 Met Cys Ser Gln Val Leu Cys Gln Glu Gln Gly Val Cys Ile Arg Lys 340 345 350 Asn Trp Asn Ser Ser Asp Tyr Leu His Leu Asn Pro Asp Asn Phe Ala 355 360 365 Ile Gln Leu Glu Lys Gly Gly Lys Phe Thr Val Arg Gly Lys Pro Thr 370 375 380 Leu Glu Asp Leu Glu Gln Phe Ser Glu Lys Phe Tyr Cys Ser Cys Tyr 385 390 395 400 Ser Thr Leu Ser Cys Lys Glu Lys Ala Asp Val Lys Asp Thr Asp Ala 405 410 415 Val Asp Val Cys Ile Ala Asp Gly Val Cys Ile Asp Ala Phe Leu Lys 420 425 430 Pro Pro Met Glu Thr Glu Glu Pro Gln Ile Phe Tyr Asn Ala Ser Pro 435 440 445 Ser Thr Leu Ser 450 ＜210＞ 21 ＜211＞ 452 ＜212＞ PRT ＜213＞ Artificial sequence <220> ＜223＞ Synthetic sequence ＜400＞ 21 Phe Arg Gly Pro Leu Leu Pro Asn Arg Pro Phe Thr Thr Val Trp Asn 1 5 10 15 Ala Pro Ser Glu Phe Cys Leu Gly Lys Phe Asp Glu Pro Leu Asp Met 20 25 30 Ser Leu Phe Ser Phe Ile Gly Ser Pro Arg Ile Asn Ala Thr Gly Gln 35 40 45 Gly Val Thr Ile Phe Tyr Val Asp Arg Leu Gly Tyr Tyr Pro Tyr Ile 50 55 60 Asp Ser Ile Thr Gly Val Thr Val Asn Gly Gly Ile Pro Gln Lys Ile 65 70 75 80 Ser Leu Gln Asp His Leu Asp Lys Ala Lys Lys Asp Ile Thr Phe Tyr 85 90 95 Met Pro Val Asp Asn Leu Gly Met Ala Val Ile Asp Trp Glu Glu Trp 100 105 110 Arg Pro Thr Trp Ala Arg Asn Trp Lys Pro Lys Asp Val Tyr Lys Asn 115 120 125 Arg Ser Ile Glu Leu Val Gln Gln Gln Asn Val Gln Leu Ser Leu Thr 130 135 140 Glu Ala Thr Glu Lys Ala Lys Gln Glu Phe Glu Lys Ala Gly Lys Asp 145 150 155 160 Phe Leu Val Glu Thr Ile Lys Leu Gly Lys Leu Leu Arg Pro Asn His 165 170 175 Leu Trp Gly Tyr Tyr Leu Phe Pro Asp Cys Tyr Asn His His Tyr Lys 180 185 190 Lys Pro Gly Tyr Asn Gly Ser Cys Phe Asn Val Glu Ile Lys Arg Asn 195 200 205 Asp Asp Leu Ser Trp Leu Trp Asn Glu Ser Thr Ala Leu Tyr Pro Ser 210 215 220 Ile Tyr Leu Asn Thr Gln Gln Ser Pro Val Ala Ala Thr Leu Tyr Val 225 230 235 240 Arg Asn Arg Val Arg Glu Ala Ile Arg Val Ser Lys Ile Pro Asp Ala 245 250 255 Lys Ser Pro Leu Pro Val Phe Ala Tyr Thr Arg Ile Val Phe Thr Asp 260 265 270 Gln Val Leu Lys Phe Leu Ser Gln Asp Glu Leu Val Tyr Thr Phe Gly 275 280 285 Glu Thr Val Ala Leu Gly Ala Ser Gly Ile Val Ile Trp Gly Thr Leu 290 295 300 Ser Ile Thr Arg Thr Lys Glu Ser Cys Gln Ala Ile Lys Glu Tyr Met 305 310 315 320 Asp Thr Thr Leu Asn Pro Tyr Ile Ile Asn Val Thr Leu Ala Ala Lys 325 330 335 Met Cys Ser Gln Val Leu Cys Gln Glu Gln Gly Val Cys Ile Arg Lys 340 345 350 Asn Trp Asn Ser Ser Asp Tyr Leu His Leu Asn Pro Asp Asn Phe Ala 355 360 365 Ile Gln Leu Glu Lys Gly Gly Lys Phe Thr Val Arg Gly Lys Pro Thr 370 375 380 Leu Glu Asp Leu Glu Gln Phe Ser Glu Lys Phe Tyr Cys Ser Cys Tyr 385 390 395 400 Ser Thr Leu Ser Cys Lys Glu Lys Ala Asp Val Lys Asp Thr Asp Ala 405 410 415 Val Asp Val Cys Ile Ala Asp Gly Val Cys Ile Asp Ala Phe Leu Lys 420 425 430 Pro Pro Met Glu Thr Glu Glu Pro Gln Ile Phe Tyr Asn Ala Ser Pro 435 440 445 Ser Thr Leu Ser 450 ＜210＞ 22 <211> 433 ＜212＞ PRT ＜213＞ Artificial sequence <220> ＜223＞ Synthetic sequence ＜400＞ 22 Phe Arg Ala Pro Pro Val Ile Pro Asn Val Pro Phe Leu Trp Ala Trp 1 5 10 15 Asn Ala Pro Ser Glu Phe Cys Leu Gly Lys Phe Asp Glu Pro Leu Asp 20 25 30 Met Ser Leu Phe Ser Phe Ile Gly Ser Pro Arg Ile Asn Ala Thr Gly 35 40 45 Gln Gly Val Thr Ile Phe Tyr Val Asp Arg Leu Gly Tyr Tyr Pro Tyr 50 55 60 Ile Asp Ser Ile Thr Gly Val Thr Val Asn Gly Gly Ile Pro Gln Lys 65 70 75 80 Ile Ser Leu Gln Asp His Leu Asp Lys Ala Lys Lys Asp Ile Thr Phe 85 90 95 Tyr Met Pro Val Asp Asn Leu Gly Met Ala Val Ile Asp Trp Glu Glu 100 105 110 Trp Arg Pro Thr Trp Ala Arg Asn Trp Lys Pro Lys Asp Val Tyr Lys 115 120 125 Asn Arg Ser Ile Glu Leu Val Gln Gln Gln Asn Val Gln Leu Ser Leu 130 135 140 Thr Glu Ala Thr Glu Lys Ala Lys Gln Glu Phe Glu Lys Ala Gly Lys 145 150 155 160 Asp Phe Leu Val Glu Thr Ile Lys Leu Gly Lys Leu Leu Arg Pro Asn 165 170 175 His Leu Trp Gly Tyr Tyr Leu Phe Pro Asp Cys Tyr Asn His His Tyr 180 185 190 Lys Lys Pro Gly Tyr Asn Gly Ser Cys Phe Asn Val Glu Ile Lys Arg 195 200 205 Asn Asp Asp Leu Ser Trp Leu Trp Asn Glu Ser Thr Ala Leu Tyr Pro 210 215 220 Ser Ile Tyr Leu Asn Thr Gln Gln Ser Pro Val Ala Ala Thr Leu Tyr 225 230 235 240 Val Arg Asn Arg Val Arg Glu Ala Ile Arg Val Ser Lys Ile Pro Asp 245 250 255 Ala Lys Ser Pro Leu Pro Val Phe Ala Tyr Thr Arg Ile Val Phe Thr 260 265 270 Asp Gln Val Leu Lys Phe Leu Ser Gln Asp Glu Leu Val Tyr Thr Phe 275 280 285 Gly Glu Thr Val Ala Leu Gly Ala Ser Gly Ile Val Ile Trp Gly Thr 290 295 300 Trp Glu Asn Thr Arg Thr Lys Glu Ser Cys Gln Ala Ile Lys Glu Tyr 305 310 315 320 Met Asp Thr Thr Leu Asn Pro Tyr Ile Ile Asn Val Thr Leu Ala Ala 325 330 335 Lys Met Cys Ser Gln Val Leu Cys Gln Glu Gln Gly Val Cys Ile Arg 340 345 350 Lys Asn Trp Asn Ser Ser Asp Tyr Leu His Leu Asn Pro Asp Asn Phe 355 360 365 Ala Ile Gln Leu Glu Lys Gly Gly Lys Phe Thr Val Arg Gly Lys Pro 370 375 380 Thr Leu Glu Asp Leu Glu Gln Phe Ser Glu Lys Phe Tyr Cys Ser Cys 385 390 395 400 Tyr Ser Thr Leu Ser Cys Lys Glu Lys Ala Asp Val Lys Asp Thr Asp 405 410 415 Ala Val Asp Val Cys Ile Ala Asp Gly Val Cys Ile Asp Ala Phe Leu 420 425 430 Lys ＜210＞ 23 ＜211＞ 431 ＜212＞ PRT ＜213＞ Artificial sequence <220> ＜223＞ Synthetic sequence ＜400＞ 23 Phe Arg Ala Pro Pro Val Ile Pro Asn Val Pro Phe Leu Trp Ala Trp 1 5 10 15 Asn Ala Pro Ser Glu Phe Cys Leu Gly Lys Phe Asp Glu Pro Leu Asp 20 25 30 Met Ser Leu Phe Ser Phe Ile Gly Ser Pro Arg Ile Asn Ala Thr Gly 35 40 45 Gln Gly Val Thr Ile Phe Tyr Val Asp Arg Leu Gly Tyr Tyr Pro Tyr 50 55 60 Ile Asp Ser Ile Thr Gly Val Thr Val Asn Gly Gly Ile Pro Gln Lys 65 70 75 80 Ile Ser Leu Gln Asp His Leu Asp Lys Ala Lys Lys Asp Ile Thr Phe 85 90 95 Tyr Met Pro Val Asp Asn Leu Gly Met Ala Val Ile Asp Trp Glu Glu 100 105 110 Trp Arg Pro Thr Trp Ala Arg Asn Trp Lys Pro Lys Asp Val Tyr Lys 115 120 125 Asn Arg Ser Ile Glu Leu Val Gln Gln Gln Asn Val Gln Leu Ser Leu 130 135 140 Thr Glu Ala Thr Glu Lys Ala Lys Gln Glu Phe Glu Lys Ala Gly Lys 145 150 155 160 Asp Phe Leu Val Glu Thr Ile Lys Leu Gly Lys Leu Leu Arg Pro Asn 165 170 175 His Leu Trp Gly Tyr Tyr Leu Phe Pro Asp Cys Tyr Asn His His Tyr 180 185 190 Lys Lys Pro Gly Tyr Asn Gly Ser Cys Phe Asn Val Glu Ile Lys Arg 195 200 205 Asn Asp Asp Leu Ser Trp Leu Trp Asn Glu Ser Thr Ala Leu Tyr Pro 210 215 220 Ser Ile Tyr Leu Asn Thr Gln Gln Ser Pro Val Ala Ala Thr Leu Tyr 225 230 235 240 Val Arg Asn Arg Val Arg Glu Ala Ile Arg Val Ser Lys Ile Pro Asp 245 250 255 Ala Lys Ser Pro Leu Pro Val Phe Ala Tyr Thr Arg Ile Val Phe Thr 260 265 270 Asp Gln Val Leu Lys Phe Leu Ser Gln Asp Glu Leu Val Tyr Thr Phe 275 280 285 Gly Glu Thr Val Ala Leu Gly Ala Ser Gly Ile Val Ile Trp Gly Thr 290 295 300 Trp Glu Asn Thr Arg Thr Lys Glu Ser Cys Gln Ala Ile Lys Glu Tyr 305 310 315 320 Met Asp Thr Thr Leu Asn Pro Tyr Ile Ile Asn Val Thr Leu Ala Ala 325 330 335 Lys Met Cys Ser Gln Val Leu Cys Gln Glu Gln Gly Val Cys Ile Arg 340 345 350 Lys Asn Trp Asn Ser Ser Asp Tyr Leu His Leu Asn Pro Asp Asn Phe 355 360 365 Ala Ile Gln Leu Glu Lys Gly Gly Lys Phe Thr Val Arg Gly Lys Pro 370 375 380 Thr Leu Glu Asp Leu Glu Gln Phe Ser Glu Lys Phe Tyr Cys Ser Cys 385 390 395 400 Tyr Ser Thr Leu Ser Cys Lys Glu Lys Ala Asp Val Lys Asp Thr Asp 405 410 415 Ala Val Asp Val Cys Ile Ala Asp Gly Val Cys Ile Asp Ala Phe 420 425 430 ＜210＞ 24 ＜211＞ 455 ＜212＞ PRT ＜213＞ Artificial sequence <220> ＜223＞ Synthetic sequence ＜400＞ 24 Leu Asn Phe Arg Ala Pro Pro Val Ile Pro Asn Val Pro Phe Leu Trp 1 5 10 15 Ala Trp Asn Ala Pro Ser Glu Phe Cys Leu Gly Lys Phe Asp Glu Pro 20 25 30 Leu Asp Met Ser Leu Phe Ser Phe Ile Gly Ser Pro Arg Ile Asn Ala 35 40 45 Thr Gly Gln Gly Val Thr Ile Phe Tyr Val Asp Arg Leu Gly Tyr Tyr 50 55 60 Pro Tyr Ile Asp Ser Ile Thr Gly Val Thr Val Asn Gly Gly Ile Pro 65 70 75 80 Gln Lys Ile Ser Leu Gln Asp His Leu Asp Lys Ala Lys Lys Lys Asp Ile 85 90 95 Thr Phe Tyr Met Pro Val Asp Asn Leu Gly Met Ala Val Ile Asp Trp 100 105 110 Glu Glu Trp Arg Pro Thr Trp Ala Arg Asn Trp Lys Pro Lys Asp Val 115 120 125 Tyr Lys Asn Arg Ser Ile Glu Leu Val Gln Gln Gln Asn Val Gln Leu 130 135 140 Ser Leu Thr Glu Ala Thr Glu Lys Ala Lys Gln Glu Phe Glu Lys Ala 145 150 155 160 Gly Lys Asp Phe Leu Val Glu Thr Ile Lys Leu Gly Lys Leu Leu Arg 165 170 175 Pro Asn His Leu Trp Gly Tyr Tyr Leu Phe Pro Asp Cys Tyr Asn His 180 185 190 His Tyr Lys Lys Pro Gly Tyr Asn Gly Ser Cys Phe Asn Val Glu Ile 195 200 205 Lys Arg Asn Asp Asp Leu Ser Trp Leu Trp Asn Glu Ser Thr Ala Leu 210 215 220 Tyr Pro Ser Ile Tyr Leu Asn Thr Gln Gln Ser Pro Val Ala Ala Thr 225 230 235 240 Leu Tyr Val Arg Asn Arg Val Arg Glu Ala Ile Arg Val Ser Lys Ile 245 250 255 Pro Asp Ala Lys Ser Pro Leu Pro Val Phe Ala Tyr Thr Arg Ile Val 260 265 270 Phe Thr Asp Gln Val Leu Lys Phe Leu Ser Gln Asp Glu Leu Val Tyr 275 280 285 Thr Phe Gly Glu Thr Val Ala Leu Gly Ala Ser Gly Ile Val Ile Trp 290 295 300 Gly Ser Trp Glu Asn Thr Arg Thr Lys Glu Ser Cys Gln Ala Ile Lys 305 310 315 320 Glu Tyr Met Asp Thr Thr Leu Asn Pro Tyr Ile Ile Asn Val Thr Leu 325 330 335 Ala Ala Lys Met Cys Ser Gln Val Leu Cys Gln Glu Gln Gly Val Cys 340 345 350 Ile Arg Lys Asn Trp Asn Ser Ser Asp Tyr Leu His Leu Asn Pro Asp 355 360 365 Asn Phe Ala Ile Gln Leu Glu Lys Gly Gly Lys Phe Thr Val Arg Gly 370 375 380 Lys Pro Thr Leu Glu Asp Leu Glu Gln Phe Ser Glu Lys Phe Tyr Cys 385 390 395 400 Ser Cys Tyr Ser Thr Leu Ser Cys Lys Glu Lys Ala Asp Val Lys Asp 405 410 415 Thr Asp Ala Val Asp Val Cys Ile Ala Asp Gly Val Cys Ile Asp Ala 420 425 430 Phe Leu Lys Pro Pro Met Glu Thr Glu Glu Pro Gln Ile Phe Tyr Asn 435 440 445 Ala Ser Pro Ser Thr Leu Ser 450 455 ＜210＞ 25 ＜211＞ 451 ＜212＞ PRT ＜213＞ Artificial sequence <220> ＜223＞ Synthetic sequence ＜400＞ 25 Ala Pro Pro Val Ile Pro Asn Val Pro Phe Leu Trp Ala Trp Asn Ala 1 5 10 15 Pro Ser Glu Phe Cys Leu Gly Lys Phe Asp Glu Pro Leu Asp Met Ser 20 25 30 Leu Phe Ser Phe Ile Gly Ser Pro Arg Ile Asn Ala Thr Gly Gln Gly 35 40 45 Val Thr Ile Phe Tyr Val Asp Arg Leu Gly Tyr Tyr Pro Tyr Ile Asp 50 55 60 Ser Ile Thr Gly Val Thr Val Asn Gly Gly Ile Pro Gln Lys Ile Ser 65 70 75 80 Leu Gln Asp His Leu Asp Lys Ala Lys Lys Asp Ile Thr Phe Tyr Met 85 90 95 Pro Val Asp Asn Leu Gly Met Ala Val Ile Asp Trp Glu Glu Trp Arg 100 105 110 Pro Thr Trp Ala Arg Asn Trp Lys Pro Lys Asp Val Tyr Lys Asn Arg 115 120 125 Ser Ile Glu Leu Val Gln Gln Gln Asn Val Gln Leu Ser Leu Thr Glu 130 135 140 Ala Thr Glu Lys Ala Lys Gln Glu Phe Glu Lys Ala Gly Lys Asp Phe 145 150 155 160 Leu Val Glu Thr Ile Lys Leu Gly Lys Leu Leu Arg Pro Asn His Leu 165 170 175 Trp Gly Tyr Tyr Leu Phe Pro Asp Cys Tyr Asn His His Tyr Lys Lys 180 185 190 Pro Gly Tyr Asn Gly Ser Cys Phe Asn Val Glu Ile Lys Arg Asn Asp 195 200 205 Asp Leu Ser Trp Leu Trp Asn Glu Ser Thr Ala Leu Tyr Pro Ser Ile 210 215 220 Tyr Leu Asn Thr Gln Gln Ser Pro Val Ala Ala Thr Leu Tyr Val Arg 225 230 235 240 Asn Arg Val Arg Glu Ala Ile Arg Val Ser Lys Ile Pro Asp Ala Lys 245 250 255 Ser Pro Leu Pro Val Phe Ala Tyr Thr Arg Ile Val Phe Thr Asp Gln 260 265 270 Val Leu Lys Phe Leu Ser Gln Asp Glu Leu Val Tyr Thr Phe Gly Glu 275 280 285 Thr Val Ala Leu Gly Ala Ser Gly Ile Val Ile Trp Gly Thr Leu Ser 290 295 300 Ile Thr Arg Thr Lys Glu Ser Cys Gln Ala Ile Lys Glu Tyr Met Asp 305 310 315 320 Thr Thr Leu Asn Pro Tyr Ile Ile Asn Val Thr Leu Ala Ala Lys Met 325 330 335 Cys Ser Gln Val Leu Cys Gln Glu Gln Gly Val Cys Ile Arg Lys Asn 340 345 350 Trp Asn Ser Ser Asp Tyr Leu His Leu Asn Pro Asp Asn Phe Ala Ile 355 360 365 Gln Leu Glu Lys Gly Gly Lys Phe Thr Val Arg Gly Lys Pro Thr Leu 370 375 380 Glu Asp Leu Glu Gln Phe Ser Glu Lys Phe Tyr Cys Ser Cys Tyr Ser 385 390 395 400 Thr Leu Ser Cys Lys Glu Lys Ala Asp Val Lys Asp Thr Asp Ala Val 405 410 415 Asp Val Cys Ile Ala Asp Gly Val Cys Ile Asp Ala Phe Leu Lys Pro 420 425 430 Pro Met Glu Thr Glu Glu Pro Gln Ile Phe Tyr Asn Ala Ser Pro Ser 435 440 445 Thr Leu Ser 450 ＜210＞ 26 ＜211＞ 449 ＜212＞ PRT ＜213＞ Artificial sequence <220> ＜223＞ Synthetic sequence ＜400＞ 26 Pro Val Ile Pro Asn Val Pro Phe Leu Trp Ala Trp Asn Ala Pro Ser 1 5 10 15 Glu Phe Cys Leu Gly Lys Phe Asp Glu Pro Leu Asp Met Ser Leu Phe 20 25 30 Ser Phe Ile Gly Ser Pro Arg Ile Asn Ala Thr Gly Gln Gly Val Thr 35 40 45 Ile Phe Tyr Val Asp Arg Leu Gly Tyr Tyr Pro Tyr Ile Asp Ser Ile 50 55 60 Thr Gly Val Thr Val Asn Gly Gly Ile Pro Gln Lys Ile Ser Leu Gln 65 70 75 80 Asp His Leu Asp Lys Ala Lys Lys Asp Ile Thr Phe Tyr Met Pro Val 85 90 95 Asp Asn Leu Gly Met Ala Val Ile Asp Trp Glu Glu Trp Arg Pro Thr 100 105 110 Trp Ala Arg Asn Trp Lys Pro Lys Asp Val Tyr Lys Asn Arg Ser Ile 115 120 125 Glu Leu Val Gln Gln Gln Asn Val Gln Leu Ser Leu Thr Glu Ala Thr 130 135 140 Glu Lys Ala Lys Gln Glu Phe Glu Lys Ala Gly Lys Asp Phe Leu Val 145 150 155 160 Glu Thr Ile Lys Leu Gly Lys Leu Leu Arg Pro Asn His Leu Trp Gly 165 170 175 Tyr Tyr Leu Phe Pro Asp Cys Tyr Asn His His Tyr Lys Lys Pro Gly 180 185 190 Tyr Asn Gly Ser Cys Phe Asn Val Glu Ile Lys Arg Asn Asp Asp Leu 195 200 205 Ser Trp Leu Trp Asn Glu Ser Thr Ala Leu Tyr Pro Ser Ile Tyr Leu 210 215 220 Asn Thr Gln Gln Ser Pro Val Ala Ala Thr Leu Tyr Val Arg Asn Arg 225 230 235 240 Val Arg Glu Ala Ile Arg Val Ser Lys Ile Pro Asp Ala Lys Ser Pro 245 250 255 Leu Pro Val Phe Ala Tyr Thr Arg Ile Val Phe Thr Asp Gln Val Leu 260 265 270 Lys Phe Leu Ser Gln Asp Glu Leu Val Tyr Thr Phe Gly Glu Thr Val 275 280 285 Ala Leu Gly Ala Ser Gly Ile Val Ile Trp Gly Thr Leu Ser Ile Thr 290 295 300 Arg Thr Lys Glu Ser Cys Gln Ala Ile Lys Glu Tyr Met Asp Thr Thr 305 310 315 320 Leu Asn Pro Tyr Ile Ile Asn Val Thr Leu Ala Ala Lys Met Cys Ser 325 330 335 Gln Val Leu Cys Gln Glu Gln Gly Val Cys Ile Arg Lys Asn Trp Asn 340 345 350 Ser Ser Asp Tyr Leu His Leu Asn Pro Asp Asn Phe Ala Ile Gln Leu 355 360 365 Glu Lys Gly Gly Lys Phe Thr Val Arg Gly Lys Pro Thr Leu Glu Asp 370 375 380 Leu Glu Gln Phe Ser Glu Lys Phe Tyr Cys Ser Cys Tyr Ser Thr Leu 385 390 395 400 Ser Cys Lys Glu Lys Ala Asp Val Lys Asp Thr Asp Ala Val Asp Val 405 410 415 Cys Ile Ala Asp Gly Val Cys Ile Asp Ala Phe Leu Lys Pro Pro Met 420 425 430 Glu Thr Glu Glu Pro Gln Ile Phe Tyr Asn Ala Ser Pro Ser Thr Leu 435 440 445 Ser ＜210＞ 27 <211> 432 ＜212＞ PRT ＜213＞ Artificial sequence <220> ＜223＞ Synthetic sequence ＜400＞ 27 Leu Asn Phe Arg Ala Pro Pro Val Ile Pro Asn Val Pro Phe Leu Trp 1 5 10 15 Ala Trp Asn Ala Pro Ser Glu Phe Cys Leu Gly Lys Phe Asp Glu Pro 20 25 30 Leu Asp Met Ser Leu Phe Ser Phe Ile Gly Ser Pro Arg Ile Asn Ala 35 40 45 Thr Gly Gln Gly Val Thr Ile Phe Tyr Val Asp Arg Leu Gly Tyr Tyr 50 55 60 Pro Tyr Ile Asp Ser Ile Thr Gly Val Thr Val Asn Gly Gly Ile Pro 65 70 75 80 Gln Lys Ile Ser Leu Gln Asp His Leu Asp Lys Ala Lys Lys Lys Asp Ile 85 90 95 Thr Phe Tyr Met Pro Val Asp Asn Leu Gly Met Ala Val Ile Asp Trp 100 105 110 Glu Glu Trp Arg Pro Thr Trp Ala Arg Asn Trp Lys Pro Lys Asp Val 115 120 125 Tyr Lys Asn Arg Ser Ile Glu Leu Val Gln Gln Gln Asn Val Gln Leu 130 135 140 Ser Leu Thr Glu Ala Thr Glu Lys Ala Lys Gln Glu Phe Glu Lys Ala 145 150 155 160 Gly Lys Asp Phe Leu Val Glu Thr Ile Lys Leu Gly Lys Leu Leu Arg 165 170 175 Pro Asn His Leu Trp Gly Tyr Tyr Leu Phe Pro Asp Cys Tyr Asn His 180 185 190 His Tyr Lys Lys Pro Gly Tyr Asn Gly Ser Cys Phe Asn Val Glu Ile 195 200 205 Lys Arg Asn Asp Asp Leu Ser Trp Leu Trp Asn Glu Ser Thr Ala Leu 210 215 220 Tyr Pro Ser Ile Tyr Leu Asn Thr Gln Gln Ser Pro Val Ala Ala Thr 225 230 235 240 Leu Tyr Val Arg Asn Arg Val Arg Glu Ala Ile Arg Val Ser Lys Ile 245 250 255 Pro Asp Ala Lys Ser Pro Leu Pro Val Phe Ala Tyr Thr Arg Ile Val 260 265 270 Phe Thr Asp Gln Val Leu Lys Phe Leu Ser Gln Asp Glu Leu Val Tyr 275 280 285 Thr Phe Gly Glu Thr Val Ala Leu Gly Ala Ser Gly Ile Val Ile Trp 290 295 300 Gly Thr Trp Glu Asn Thr Arg Thr Lys Glu Ser Cys Gln Ala Ile Lys 305 310 315 320 Glu Tyr Met Asp Thr Thr Leu Asn Pro Tyr Ile Ile Asn Val Thr Leu 325 330 335 Ala Ala Lys Met Cys Ser Gln Val Leu Cys Gln Glu Gln Gly Val Cys 340 345 350 Ile Arg Lys Asn Trp Asn Ser Ser Asp Tyr Leu His Leu Asn Pro Asp 355 360 365 Asn Phe Ala Ile Gln Leu Glu Lys Gly Gly Lys Phe Thr Val Arg Gly 370 375 380 Lys Pro Thr Leu Glu Asp Leu Glu Gln Phe Ser Glu Lys Phe Tyr Cys 385 390 395 400 Ser Cys Tyr Ser Thr Leu Ser Cys Lys Glu Lys Ala Asp Val Lys Asp 405 410 415 Thr Asp Ala Val Asp Val Cys Ile Ala Asp Gly Val Cys Ile Asp Ala 420 425 430 ＜210＞ 28 ＜211＞ 429 ＜212＞ PRT ＜213＞ Artificial sequence <220> ＜223＞ Synthetic sequence ＜400＞ 28 Leu Asn Phe Arg Ala Pro Pro Val Ile Pro Asn Val Pro Phe Leu Trp 1 5 10 15 Ala Trp Asn Ala Pro Ser Glu Phe Cys Leu Gly Lys Phe Asp Glu Pro 20 25 30 Leu Asp Met Ser Leu Phe Ser Phe Ile Gly Ser Pro Arg Ile Asn Ala 35 40 45 Thr Gly Gln Gly Val Thr Ile Phe Tyr Val Asp Arg Leu Gly Tyr Tyr 50 55 60 Pro Tyr Ile Asp Ser Ile Thr Gly Val Thr Val Asn Gly Gly Ile Pro 65 70 75 80 Gln Lys Ile Ser Leu Gln Asp His Leu Asp Lys Ala Lys Lys Lys Asp Ile 85 90 95 Thr Phe Tyr Met Pro Val Asp Asn Leu Gly Met Ala Val Ile Asp Trp 100 105 110 Glu Glu Trp Arg Pro Thr Trp Ala Arg Asn Trp Lys Pro Lys Asp Val 115 120 125 Tyr Lys Asn Arg Ser Ile Glu Leu Val Gln Gln Gln Asn Val Gln Leu 130 135 140 Ser Leu Thr Glu Ala Thr Glu Lys Ala Lys Gln Glu Phe Glu Lys Ala 145 150 155 160 Gly Lys Asp Phe Leu Val Glu Thr Ile Lys Leu Gly Lys Leu Leu Arg 165 170 175 Pro Asn His Leu Trp Gly Tyr Tyr Leu Phe Pro Asp Cys Tyr Asn His 180 185 190 His Tyr Lys Lys Pro Gly Tyr Asn Gly Ser Cys Phe Asn Val Glu Ile 195 200 205 Lys Arg Asn Asp Asp Leu Ser Trp Leu Trp Asn Glu Ser Thr Ala Leu 210 215 220 Tyr Pro Ser Ile Tyr Leu Asn Thr Gln Gln Ser Pro Val Ala Ala Thr 225 230 235 240 Leu Tyr Val Arg Asn Arg Val Arg Glu Ala Ile Arg Val Ser Lys Ile 245 250 255 Pro Asp Ala Lys Ser Pro Leu Pro Val Phe Ala Tyr Thr Arg Ile Val 260 265 270 Phe Thr Asp Gln Val Leu Lys Phe Leu Ser Gln Asp Glu Leu Val Tyr 275 280 285 Thr Phe Gly Glu Thr Val Ala Leu Gly Ala Ser Gly Ile Val Ile Trp 290 295 300 Gly Thr Trp Glu Asn Thr Arg Thr Lys Glu Ser Cys Gln Ala Ile Lys 305 310 315 320 Glu Tyr Met Asp Thr Thr Leu Asn Pro Tyr Ile Ile Asn Val Thr Leu 325 330 335 Ala Ala Lys Met Cys Ser Gln Val Leu Cys Gln Glu Gln Gly Val Cys 340 345 350 Ile Arg Lys Asn Trp Asn Ser Ser Asp Tyr Leu His Leu Asn Pro Asp 355 360 365 Asn Phe Ala Ile Gln Leu Glu Lys Gly Gly Lys Phe Thr Val Arg Gly 370 375 380 Lys Pro Thr Leu Glu Asp Leu Glu Gln Phe Ser Glu Lys Phe Tyr Cys 385 390 395 400 Ser Cys Tyr Ser Thr Leu Ser Cys Lys Glu Lys Ala Asp Val Lys Asp 405 410 415 Thr Asp Ala Val Asp Val Cys Ile Ala Asp Gly Val Cys 420 425 ＜210＞ 29 ＜211＞ 426 ＜212＞ PRT ＜213＞ Artificial sequence <220> ＜223＞ Synthetic sequence ＜400＞ 29 Leu Asn Phe Arg Ala Pro Pro Val Ile Pro Asn Val Pro Phe Leu Trp 1 5 10 15 Ala Trp Asn Ala Pro Ser Glu Phe Cys Leu Gly Lys Phe Asp Glu Pro 20 25 30 Leu Asp Met Ser Leu Phe Ser Phe Ile Gly Ser Pro Arg Ile Asn Ala 35 40 45 Thr Gly Gln Gly Val Thr Ile Phe Tyr Val Asp Arg Leu Gly Tyr Tyr 50 55 60 Pro Tyr Ile Asp Ser Ile Thr Gly Val Thr Val Asn Gly Gly Ile Pro 65 70 75 80 Gln Lys Ile Ser Leu Gln Asp His Leu Asp Lys Ala Lys Lys Lys Asp Ile 85 90 95 Thr Phe Tyr Met Pro Val Asp Asn Leu Gly Met Ala Val Ile Asp Trp 100 105 110 Glu Glu Trp Arg Pro Thr Trp Ala Arg Asn Trp Lys Pro Lys Asp Val 115 120 125 Tyr Lys Asn Arg Ser Ile Glu Leu Val Gln Gln Gln Asn Val Gln Leu 130 135 140 Ser Leu Thr Glu Ala Thr Glu Lys Ala Lys Gln Glu Phe Glu Lys Ala 145 150 155 160 Gly Lys Asp Phe Leu Val Glu Thr Ile Lys Leu Gly Lys Leu Leu Arg 165 170 175 Pro Asn His Leu Trp Gly Tyr Tyr Leu Phe Pro Asp Cys Tyr Asn His 180 185 190 His Tyr Lys Lys Pro Gly Tyr Asn Gly Ser Cys Phe Asn Val Glu Ile 195 200 205 Lys Arg Asn Asp Asp Leu Ser Trp Leu Trp Asn Glu Ser Thr Ala Leu 210 215 220 Tyr Pro Ser Ile Tyr Leu Asn Thr Gln Gln Ser Pro Val Ala Ala Thr 225 230 235 240 Leu Tyr Val Arg Asn Arg Val Arg Glu Ala Ile Arg Val Ser Lys Ile 245 250 255 Pro Asp Ala Lys Ser Pro Leu Pro Val Phe Ala Tyr Thr Arg Ile Val 260 265 270 Phe Thr Asp Gln Val Leu Lys Phe Leu Ser Gln Asp Glu Leu Val Tyr 275 280 285 Thr Phe Gly Glu Thr Val Ala Leu Gly Ala Ser Gly Ile Val Ile Trp 290 295 300 Gly Thr Trp Glu Asn Thr Arg Thr Lys Glu Ser Cys Gln Ala Ile Lys 305 310 315 320 Glu Tyr Met Asp Thr Thr Leu Asn Pro Tyr Ile Ile Asn Val Thr Leu 325 330 335 Ala Ala Lys Met Cys Ser Gln Val Leu Cys Gln Glu Gln Gly Val Cys 340 345 350 Ile Arg Lys Asn Trp Asn Ser Ser Asp Tyr Leu His Leu Asn Pro Asp 355 360 365 Asn Phe Ala Ile Gln Leu Glu Lys Gly Gly Lys Phe Thr Val Arg Gly 370 375 380 Lys Pro Thr Leu Glu Asp Leu Glu Gln Phe Ser Glu Lys Phe Tyr Cys 385 390 395 400 Ser Cys Tyr Ser Thr Leu Ser Cys Lys Glu Lys Ala Asp Val Lys Asp 405 410 415 Thr Asp Ala Val Asp Val Cys Ile Ala Asp 420 425 ＜210＞ 30 <211> 423 ＜212＞ PRT ＜213＞ Artificial sequence <220> ＜223＞ Synthetic sequence ＜400＞ 30 Leu Asn Phe Arg Ala Pro Pro Val Ile Pro Asn Val Pro Phe Leu Trp 1 5 10 15 Ala Trp Asn Ala Pro Ser Glu Phe Cys Leu Gly Lys Phe Asp Glu Pro 20 25 30 Leu Asp Met Ser Leu Phe Ser Phe Ile Gly Ser Pro Arg Ile Asn Ala 35 40 45 Thr Gly Gln Gly Val Thr Ile Phe Tyr Val Asp Arg Leu Gly Tyr Tyr 50 55 60 Pro Tyr Ile Asp Ser Ile Thr Gly Val Thr Val Asn Gly Gly Ile Pro 65 70 75 80 Gln Lys Ile Ser Leu Gln Asp His Leu Asp Lys Ala Lys Lys Lys Asp Ile 85 90 95 Thr Phe Tyr Met Pro Val Asp Asn Leu Gly Met Ala Val Ile Asp Trp 100 105 110 Glu Glu Trp Arg Pro Thr Trp Ala Arg Asn Trp Lys Pro Lys Asp Val 115 120 125 Tyr Lys Asn Arg Ser Ile Glu Leu Val Gln Gln Gln Asn Val Gln Leu 130 135 140 Ser Leu Thr Glu Ala Thr Glu Lys Ala Lys Gln Glu Phe Glu Lys Ala 145 150 155 160 Gly Lys Asp Phe Leu Val Glu Thr Ile Lys Leu Gly Lys Leu Leu Arg 165 170 175 Pro Asn His Leu Trp Gly Tyr Tyr Leu Phe Pro Asp Cys Tyr Asn His 180 185 190 His Tyr Lys Lys Pro Gly Tyr Asn Gly Ser Cys Phe Asn Val Glu Ile 195 200 205 Lys Arg Asn Asp Asp Leu Ser Trp Leu Trp Asn Glu Ser Thr Ala Leu 210 215 220 Tyr Pro Ser Ile Tyr Leu Asn Thr Gln Gln Ser Pro Val Ala Ala Thr 225 230 235 240 Leu Tyr Val Arg Asn Arg Val Arg Glu Ala Ile Arg Val Ser Lys Ile 245 250 255 Pro Asp Ala Lys Ser Pro Leu Pro Val Phe Ala Tyr Thr Arg Ile Val 260 265 270 Phe Thr Asp Gln Val Leu Lys Phe Leu Ser Gln Asp Glu Leu Val Tyr 275 280 285 Thr Phe Gly Glu Thr Val Ala Leu Gly Ala Ser Gly Ile Val Ile Trp 290 295 300 Gly Thr Trp Glu Asn Thr Arg Thr Lys Glu Ser Cys Gln Ala Ile Lys 305 310 315 320 Glu Tyr Met Asp Thr Thr Leu Asn Pro Tyr Ile Ile Asn Val Thr Leu 325 330 335 Ala Ala Lys Met Cys Ser Gln Val Leu Cys Gln Glu Gln Gly Val Cys 340 345 350 Ile Arg Lys Asn Trp Asn Ser Ser Asp Tyr Leu His Leu Asn Pro Asp 355 360 365 Asn Phe Ala Ile Gln Leu Glu Lys Gly Gly Lys Phe Thr Val Arg Gly 370 375 380 Lys Pro Thr Leu Glu Asp Leu Glu Gln Phe Ser Glu Lys Phe Tyr Cys 385 390 395 400 Ser Cys Tyr Ser Thr Leu Ser Cys Lys Glu Lys Ala Asp Val Lys Asp 405 410 415 Thr Asp Ala Val Asp Val Cys 420 ＜210＞ 31 ＜211＞ 420 ＜212＞ PRT ＜213＞ Artificial sequence <220> ＜223＞ Synthetic sequence ＜400＞ 31 Leu Asn Phe Arg Ala Pro Pro Val Ile Pro Asn Val Pro Phe Leu Trp 1 5 10 15 Ala Trp Asn Ala Pro Ser Glu Phe Cys Leu Gly Lys Phe Asp Glu Pro 20 25 30 Leu Asp Met Ser Leu Phe Ser Phe Ile Gly Ser Pro Arg Ile Asn Ala 35 40 45 Thr Gly Gln Gly Val Thr Ile Phe Tyr Val Asp Arg Leu Gly Tyr Tyr 50 55 60 Pro Tyr Ile Asp Ser Ile Thr Gly Val Thr Val Asn Gly Gly Ile Pro 65 70 75 80 Gln Lys Ile Ser Leu Gln Asp His Leu Asp Lys Ala Lys Lys Lys Asp Ile 85 90 95 Thr Phe Tyr Met Pro Val Asp Asn Leu Gly Met Ala Val Ile Asp Trp 100 105 110 Glu Glu Trp Arg Pro Thr Trp Ala Arg Asn Trp Lys Pro Lys Asp Val 115 120 125 Tyr Lys Asn Arg Ser Ile Glu Leu Val Gln Gln Gln Asn Val Gln Leu 130 135 140 Ser Leu Thr Glu Ala Thr Glu Lys Ala Lys Gln Glu Phe Glu Lys Ala 145 150 155 160 Gly Lys Asp Phe Leu Val Glu Thr Ile Lys Leu Gly Lys Leu Leu Arg 165 170 175 Pro Asn His Leu Trp Gly Tyr Tyr Leu Phe Pro Asp Cys Tyr Asn His 180 185 190 His Tyr Lys Lys Pro Gly Tyr Asn Gly Ser Cys Phe Asn Val Glu Ile 195 200 205 Lys Arg Asn Asp Asp Leu Ser Trp Leu Trp Asn Glu Ser Thr Ala Leu 210 215 220 Tyr Pro Ser Ile Tyr Leu Asn Thr Gln Gln Ser Pro Val Ala Ala Thr 225 230 235 240 Leu Tyr Val Arg Asn Arg Val Arg Glu Ala Ile Arg Val Ser Lys Ile 245 250 255 Pro Asp Ala Lys Ser Pro Leu Pro Val Phe Ala Tyr Thr Arg Ile Val 260 265 270 Phe Thr Asp Gln Val Leu Lys Phe Leu Ser Gln Asp Glu Leu Val Tyr 275 280 285 Thr Phe Gly Glu Thr Val Ala Leu Gly Ala Ser Gly Ile Val Ile Trp 290 295 300 Gly Thr Trp Glu Asn Thr Arg Thr Lys Glu Ser Cys Gln Ala Ile Lys 305 310 315 320 Glu Tyr Met Asp Thr Thr Leu Asn Pro Tyr Ile Ile Asn Val Thr Leu 325 330 335 Ala Ala Lys Met Cys Ser Gln Val Leu Cys Gln Glu Gln Gly Val Cys 340 345 350 Ile Arg Lys Asn Trp Asn Ser Ser Asp Tyr Leu His Leu Asn Pro Asp 355 360 365 Asn Phe Ala Ile Gln Leu Glu Lys Gly Gly Lys Phe Thr Val Arg Gly 370 375 380 Lys Pro Thr Leu Glu Asp Leu Glu Gln Phe Ser Glu Lys Phe Tyr Cys 385 390 395 400 Ser Cys Tyr Ser Thr Leu Ser Cys Lys Glu Lys Ala Asp Val Lys Asp 405 410 415 Thr Asp Ala Val 420 ＜210＞ 32 <211> 433 ＜212＞ PRT ＜213＞ Artificial sequence <220> ＜223＞ Synthetic sequence ＜400＞ 32 Phe Arg Ala Pro Pro Val Ile Pro Asn Val Pro Phe Leu Trp Ala Trp 1 5 10 15 Asn Ala Pro Ser Glu Phe Cys Leu Gly Lys Phe Asp Glu Pro Leu Asp 20 25 30 Met Ser Leu Phe Ser Phe Ile Gly Ser Pro Arg Ile Asn Ala Thr Gly 35 40 45 Gln Gly Val Thr Ile Phe Tyr Val Asp Arg Leu Gly Tyr Tyr Pro Tyr 50 55 60 Ile Asp Ser Ile Thr Gly Val Thr Val Asn Gly Gly Ile Pro Gln Lys 65 70 75 80 Ile Ser Leu Gln Asp His Leu Asp Lys Ala Lys Lys Asp Ile Thr Phe 85 90 95 Tyr Met Pro Val Asp Asn Leu Gly Met Ala Val Ile Asp Trp Glu Glu 100 105 110 Trp Arg Pro Thr Trp Ala Arg Asn Trp Lys Pro Lys Asp Val Tyr Lys 115 120 125 Asn Arg Ser Ile Glu Leu Val Gln Gln Gln Asn Val Gln Leu Ser Leu 130 135 140 Thr Glu Ala Thr Glu Lys Ala Lys Gln Glu Phe Glu Lys Ala Gly Lys 145 150 155 160 Asp Phe Leu Val Glu Thr Ile Lys Leu Gly Lys Leu Leu Arg Pro Asn 165 170 175 His Leu Trp Gly Tyr Tyr Leu Phe Pro Asp Cys Tyr Asn His His Tyr 180 185 190 Lys Lys Pro Gly Tyr Asn Gly Ser Cys Phe Asn Val Glu Ile Lys Arg 195 200 205 Asn Asp Asp Leu Ser Trp Leu Trp Asn Glu Ser Thr Ala Leu Tyr Pro 210 215 220 Ser Ile Tyr Leu Asn Thr Gln Gln Ser Pro Val Ala Ala Thr Leu Tyr 225 230 235 240 Val Arg Asn Arg Val Arg Glu Ala Ile Arg Val Ser Lys Ile Pro Asp 245 250 255 Ala Lys Ser Pro Leu Pro Val Phe Ala Tyr Thr Arg Ile Val Phe Thr 260 265 270 Asp Gln Val Leu Lys Phe Leu Ser Gln Asp Glu Leu Val Tyr Thr Phe 275 280 285 Gly Glu Thr Val Ala Leu Gly Ala Ser Gly Ile Val Ile Trp Gly Ser 290 295 300 Trp Glu Asn Thr Arg Thr Lys Glu Ser Cys Gln Ala Ile Lys Glu Tyr 305 310 315 320 Met Asp Thr Thr Leu Asn Pro Tyr Ile Ile Asn Val Thr Leu Ala Ala 325 330 335 Lys Met Cys Ser Gln Val Leu Cys Gln Glu Gln Gly Val Cys Ile Arg 340 345 350 Lys Asn Trp Asn Ser Ser Asp Tyr Leu His Leu Asn Pro Asp Asn Phe 355 360 365 Ala Ile Gln Leu Glu Lys Gly Gly Lys Phe Thr Val Arg Gly Lys Pro 370 375 380 Thr Leu Glu Asp Leu Glu Gln Phe Ser Glu Lys Phe Tyr Cys Ser Cys 385 390 395 400 Tyr Ser Thr Leu Ser Cys Lys Glu Lys Ala Asp Val Lys Asp Thr Asp 405 410 415 Ala Val Asp Val Cys Ile Ala Asp Gly Val Cys Ile Asp Ala Phe Leu 420 425 430 Lys ＜210＞ 33 ＜211＞ 435 ＜212＞ PRT ＜213＞ Artificial sequence <220> ＜223＞ Synthetic sequence ＜400＞ 33 Phe Arg Ala Pro Pro Val Ile Pro Asn Val Pro Phe Leu Trp Ala Trp 1 5 10 15 Asn Ala Pro Ser Glu Phe Cys Leu Gly Lys Phe Asp Glu Pro Leu Asp 20 25 30 Met Ser Leu Phe Ser Phe Ile Gly Ser Pro Arg Ile Asn Ala Thr Gly 35 40 45 Gln Gly Val Thr Ile Phe Tyr Val Asp Arg Leu Gly Tyr Tyr Pro Tyr 50 55 60 Ile Asp Ser Ile Thr Gly Val Thr Val Asn Gly Gly Ile Pro Gln Lys 65 70 75 80 Ile Ser Leu Gln Asp His Leu Asp Lys Ala Lys Lys Asp Ile Thr Phe 85 90 95 Tyr Met Pro Val Asp Asn Leu Gly Met Ala Val Ile Asp Trp Glu Glu 100 105 110 Trp Arg Pro Thr Trp Ala Arg Asn Trp Lys Pro Lys Asp Val Tyr Lys 115 120 125 Asn Arg Ser Ile Glu Leu Val Gln Gln Gln Asn Val Gln Leu Ser Leu 130 135 140 Thr Glu Ala Thr Glu Lys Ala Lys Gln Glu Phe Glu Lys Ala Gly Lys 145 150 155 160 Asp Phe Leu Val Glu Thr Ile Lys Leu Gly Lys Leu Leu Arg Pro Asn 165 170 175 His Leu Trp Gly Tyr Tyr Leu Phe Pro Asp Cys Tyr Asn His His Tyr 180 185 190 Lys Lys Pro Gly Tyr Asn Gly Ser Cys Phe Asn Val Glu Ile Lys Arg 195 200 205 Asn Asp Asp Leu Ser Trp Leu Trp Asn Glu Ser Thr Ala Leu Tyr Pro 210 215 220 Ser Ile Tyr Leu Asn Thr Gln Gln Ser Pro Val Ala Ala Thr Leu Tyr 225 230 235 240 Val Arg Asn Arg Val Arg Glu Ala Ile Arg Val Ser Lys Ile Pro Asp 245 250 255 Ala Lys Ser Pro Leu Pro Val Phe Ala Tyr Thr Arg Ile Val Phe Thr 260 265 270 Asp Gln Val Leu Lys Phe Leu Ser Gln Asp Glu Leu Val Tyr Thr Phe 275 280 285 Gly Glu Thr Val Ala Leu Gly Ala Ser Gly Ile Val Ile Trp Gly Thr 290 295 300 Trp Glu Asn Thr Arg Thr Lys Glu Ser Cys Gln Ala Ile Lys Glu Tyr 305 310 315 320 Met Asp Thr Thr Leu Asn Pro Tyr Ile Ile Asn Val Thr Leu Ala Ala 325 330 335 Lys Met Cys Ser Gln Val Leu Cys Gln Glu Gln Gly Val Cys Ile Arg 340 345 350 Lys Asn Trp Asn Ser Ser Asp Tyr Leu His Leu Asn Pro Asp Asn Phe 355 360 365 Ala Ile Gln Leu Glu Lys Gly Gly Lys Phe Thr Val Arg Gly Lys Pro 370 375 380 Thr Leu Glu Asp Leu Glu Gln Phe Ser Glu Lys Phe Tyr Cys Ser Cys 385 390 395 400 Tyr Ser Thr Leu Ser Cys Lys Glu Lys Ala Asp Val Lys Asp Thr Asp 405 410 415 Ala Val Asp Val Cys Ile Ala Asp Gly Val Cys Ile Asp Ala Phe Leu 420 425 430 Lys Pro Pro 435 ＜210＞ 34 ＜211＞ 436 ＜212＞ PRT ＜213＞ Artificial sequence <220> ＜223＞ Synthetic sequence ＜400＞ 34 Phe Arg Ala Pro Pro Val Ile Pro Asn Val Pro Phe Leu Trp Ala Trp 1 5 10 15 Asn Ala Pro Ser Glu Phe Cys Leu Gly Lys Phe Asp Glu Pro Leu Asp 20 25 30 Met Ser Leu Phe Ser Phe Ile Gly Ser Pro Arg Ile Asn Ala Thr Gly 35 40 45 Gln Gly Val Thr Ile Phe Tyr Val Asp Arg Leu Gly Tyr Tyr Pro Tyr 50 55 60 Ile Asp Ser Ile Thr Gly Val Thr Val Asn Gly Gly Ile Pro Gln Lys 65 70 75 80 Ile Ser Leu Gln Asp His Leu Asp Lys Ala Lys Lys Asp Ile Thr Phe 85 90 95 Tyr Met Pro Val Asp Asn Leu Gly Met Ala Val Ile Asp Trp Glu Glu 100 105 110 Trp Arg Pro Thr Trp Ala Arg Asn Trp Lys Pro Lys Asp Val Tyr Lys 115 120 125 Asn Arg Ser Ile Glu Leu Val Gln Gln Gln Asn Val Gln Leu Ser Leu 130 135 140 Thr Glu Ala Thr Glu Lys Ala Lys Gln Glu Phe Glu Lys Ala Gly Lys 145 150 155 160 Asp Phe Leu Val Glu Thr Ile Lys Leu Gly Lys Leu Leu Arg Pro Asn 165 170 175 His Leu Trp Gly Tyr Tyr Leu Phe Pro Asp Cys Tyr Asn His His Tyr 180 185 190 Lys Lys Pro Gly Tyr Asn Gly Ser Cys Phe Asn Val Glu Ile Lys Arg 195 200 205 Asn Asp Asp Leu Ser Trp Leu Trp Asn Glu Ser Thr Ala Leu Tyr Pro 210 215 220 Ser Ile Tyr Leu Asn Thr Gln Gln Ser Pro Val Ala Ala Thr Leu Tyr 225 230 235 240 Val Arg Asn Arg Val Arg Glu Ala Ile Arg Val Ser Lys Ile Pro Asp 245 250 255 Ala Lys Ser Pro Leu Pro Val Phe Ala Tyr Thr Arg Ile Val Phe Thr 260 265 270 Asp Gln Val Leu Lys Phe Leu Ser Gln Asp Glu Leu Val Tyr Thr Phe 275 280 285 Gly Glu Thr Val Ala Leu Gly Ala Ser Gly Ile Val Ile Trp Gly Thr 290 295 300 Trp Glu Asn Thr Arg Thr Lys Glu Ser Cys Gln Ala Ile Lys Glu Tyr 305 310 315 320 Met Asp Thr Thr Leu Asn Pro Tyr Ile Ile Asn Val Thr Leu Ala Ala 325 330 335 Lys Met Cys Ser Gln Val Leu Cys Gln Glu Gln Gly Val Cys Ile Arg 340 345 350 Lys Asn Trp Asn Ser Ser Asp Tyr Leu His Leu Asn Pro Asp Asn Phe 355 360 365 Ala Ile Gln Leu Glu Lys Gly Gly Lys Phe Thr Val Arg Gly Lys Pro 370 375 380 Thr Leu Glu Asp Leu Glu Gln Phe Ser Glu Lys Phe Tyr Cys Ser Cys 385 390 395 400 Tyr Ser Thr Leu Ser Cys Lys Glu Lys Ala Asp Val Lys Asp Thr Asp 405 410 415 Ala Val Asp Val Cys Ile Ala Asp Gly Val Cys Ile Asp Ala Phe Leu 420 425 430 Lys Pro Pro Met 435 ＜210＞ 35 ＜211＞ 437 ＜212＞ PRT ＜213＞ Artificial sequence <220> ＜223＞ Synthetic sequence ＜400＞ 35 Phe Arg Ala Pro Pro Val Ile Pro Asn Val Pro Phe Leu Trp Ala Trp 1 5 10 15 Asn Ala Pro Ser Glu Phe Cys Leu Gly Lys Phe Asp Glu Pro Leu Asp 20 25 30 Met Ser Leu Phe Ser Phe Ile Gly Ser Pro Arg Ile Asn Ala Thr Gly 35 40 45 Gln Gly Val Thr Ile Phe Tyr Val Asp Arg Leu Gly Tyr Tyr Pro Tyr 50 55 60 Ile Asp Ser Ile Thr Gly Val Thr Val Asn Gly Gly Ile Pro Gln Lys 65 70 75 80 Ile Ser Leu Gln Asp His Leu Asp Lys Ala Lys Lys Asp Ile Thr Phe 85 90 95 Tyr Met Pro Val Asp Asn Leu Gly Met Ala Val Ile Asp Trp Glu Glu 100 105 110 Trp Arg Pro Thr Trp Ala Arg Asn Trp Lys Pro Lys Asp Val Tyr Lys 115 120 125 Asn Arg Ser Ile Glu Leu Val Gln Gln Gln Asn Val Gln Leu Ser Leu 130 135 140 Thr Glu Ala Thr Glu Lys Ala Lys Gln Glu Phe Glu Lys Ala Gly Lys 145 150 155 160 Asp Phe Leu Val Glu Thr Ile Lys Leu Gly Lys Leu Leu Arg Pro Asn 165 170 175 His Leu Trp Gly Tyr Tyr Leu Phe Pro Asp Cys Tyr Asn His His Tyr 180 185 190 Lys Lys Pro Gly Tyr Asn Gly Ser Cys Phe Asn Val Glu Ile Lys Arg 195 200 205 Asn Asp Asp Leu Ser Trp Leu Trp Asn Glu Ser Thr Ala Leu Tyr Pro 210 215 220 Ser Ile Tyr Leu Asn Thr Gln Gln Ser Pro Val Ala Ala Thr Leu Tyr 225 230 235 240 Val Arg Asn Arg Val Arg Glu Ala Ile Arg Val Ser Lys Ile Pro Asp 245 250 255 Ala Lys Ser Pro Leu Pro Val Phe Ala Tyr Thr Arg Ile Val Phe Thr 260 265 270 Asp Gln Val Leu Lys Phe Leu Ser Gln Asp Glu Leu Val Tyr Thr Phe 275 280 285 Gly Glu Thr Val Ala Leu Gly Ala Ser Gly Ile Val Ile Trp Gly Thr 290 295 300 Trp Glu Asn Thr Arg Thr Lys Glu Ser Cys Gln Ala Ile Lys Glu Tyr 305 310 315 320 Met Asp Thr Thr Leu Asn Pro Tyr Ile Ile Asn Val Thr Leu Ala Ala 325 330 335 Lys Met Cys Ser Gln Val Leu Cys Gln Glu Gln Gly Val Cys Ile Arg 340 345 350 Lys Asn Trp Asn Ser Ser Asp Tyr Leu His Leu Asn Pro Asp Asn Phe 355 360 365 Ala Ile Gln Leu Glu Lys Gly Gly Lys Phe Thr Val Arg Gly Lys Pro 370 375 380 Thr Leu Glu Asp Leu Glu Gln Phe Ser Glu Lys Phe Tyr Cys Ser Cys 385 390 395 400 Tyr Ser Thr Leu Ser Cys Lys Glu Lys Ala Asp Val Lys Asp Thr Asp 405 410 415 Ala Val Asp Val Cys Ile Ala Asp Gly Val Cys Ile Asp Ala Phe Leu 420 425 430 Lys Pro Pro Met Glu 435 ＜210＞ 36 ＜211＞ 438 ＜212＞ PRT ＜213＞ Artificial sequence <220> ＜223＞ Synthetic sequence ＜400＞ 36 Phe Arg Ala Pro Pro Val Ile Pro Asn Val Pro Phe Leu Trp Ala Trp 1 5 10 15 Asn Ala Pro Ser Glu Phe Cys Leu Gly Lys Phe Asp Glu Pro Leu Asp 20 25 30 Met Ser Leu Phe Ser Phe Ile Gly Ser Pro Arg Ile Asn Ala Thr Gly 35 40 45 Gln Gly Val Thr Ile Phe Tyr Val Asp Arg Leu Gly Tyr Tyr Pro Tyr 50 55 60 Ile Asp Ser Ile Thr Gly Val Thr Val Asn Gly Gly Ile Pro Gln Lys 65 70 75 80 Ile Ser Leu Gln Asp His Leu Asp Lys Ala Lys Lys Asp Ile Thr Phe 85 90 95 Tyr Met Pro Val Asp Asn Leu Gly Met Ala Val Ile Asp Trp Glu Glu 100 105 110 Trp Arg Pro Thr Trp Ala Arg Asn Trp Lys Pro Lys Asp Val Tyr Lys 115 120 125 Asn Arg Ser Ile Glu Leu Val Gln Gln Gln Asn Val Gln Leu Ser Leu 130 135 140 Thr Glu Ala Thr Glu Lys Ala Lys Gln Glu Phe Glu Lys Ala Gly Lys 145 150 155 160 Asp Phe Leu Val Glu Thr Ile Lys Leu Gly Lys Leu Leu Arg Pro Asn 165 170 175 His Leu Trp Gly Tyr Tyr Leu Phe Pro Asp Cys Tyr Asn His His Tyr 180 185 190 Lys Lys Pro Gly Tyr Asn Gly Ser Cys Phe Asn Val Glu Ile Lys Arg 195 200 205 Asn Asp Asp Leu Ser Trp Leu Trp Asn Glu Ser Thr Ala Leu Tyr Pro 210 215 220 Ser Ile Tyr Leu Asn Thr Gln Gln Ser Pro Val Ala Ala Thr Leu Tyr 225 230 235 240 Val Arg Asn Arg Val Arg Glu Ala Ile Arg Val Ser Lys Ile Pro Asp 245 250 255 Ala Lys Ser Pro Leu Pro Val Phe Ala Tyr Thr Arg Ile Val Phe Thr 260 265 270 Asp Gln Val Leu Lys Phe Leu Ser Gln Asp Glu Leu Val Tyr Thr Phe 275 280 285 Gly Glu Thr Val Ala Leu Gly Ala Ser Gly Ile Val Ile Trp Gly Thr 290 295 300 Trp Glu Asn Thr Arg Thr Lys Glu Ser Cys Gln Ala Ile Lys Glu Tyr 305 310 315 320 Met Asp Thr Thr Leu Asn Pro Tyr Ile Ile Asn Val Thr Leu Ala Ala 325 330 335 Lys Met Cys Ser Gln Val Leu Cys Gln Glu Gln Gly Val Cys Ile Arg 340 345 350 Lys Asn Trp Asn Ser Ser Asp Tyr Leu His Leu Asn Pro Asp Asn Phe 355 360 365 Ala Ile Gln Leu Glu Lys Gly Gly Lys Phe Thr Val Arg Gly Lys Pro 370 375 380 Thr Leu Glu Asp Leu Glu Gln Phe Ser Glu Lys Phe Tyr Cys Ser Cys 385 390 395 400 Tyr Ser Thr Leu Ser Cys Lys Glu Lys Ala Asp Val Lys Asp Thr Asp 405 410 415 Ala Val Asp Val Cys Ile Ala Asp Gly Val Cys Ile Asp Ala Phe Leu 420 425 430 Lys Pro Pro Met Glu Thr 435 ＜210＞ 37 ＜211＞ 439 ＜212＞ PRT ＜213＞ Artificial sequence <220> ＜223＞ Synthetic sequence ＜400＞ 37 Phe Arg Ala Pro Pro Val Ile Pro Asn Val Pro Phe Leu Trp Ala Trp 1 5 10 15 Asn Ala Pro Ser Glu Phe Cys Leu Gly Lys Phe Asp Glu Pro Leu Asp 20 25 30 Met Ser Leu Phe Ser Phe Ile Gly Ser Pro Arg Ile Asn Ala Thr Gly 35 40 45 Gln Gly Val Thr Ile Phe Tyr Val Asp Arg Leu Gly Tyr Tyr Pro Tyr 50 55 60 Ile Asp Ser Ile Thr Gly Val Thr Val Asn Gly Gly Ile Pro Gln Lys 65 70 75 80 Ile Ser Leu Gln Asp His Leu Asp Lys Ala Lys Lys Asp Ile Thr Phe 85 90 95 Tyr Met Pro Val Asp Asn Leu Gly Met Ala Val Ile Asp Trp Glu Glu 100 105 110 Trp Arg Pro Thr Trp Ala Arg Asn Trp Lys Pro Lys Asp Val Tyr Lys 115 120 125 Asn Arg Ser Ile Glu Leu Val Gln Gln Gln Asn Val Gln Leu Ser Leu 130 135 140 Thr Glu Ala Thr Glu Lys Ala Lys Gln Glu Phe Glu Lys Ala Gly Lys 145 150 155 160 Asp Phe Leu Val Glu Thr Ile Lys Leu Gly Lys Leu Leu Arg Pro Asn 165 170 175 His Leu Trp Gly Tyr Tyr Leu Phe Pro Asp Cys Tyr Asn His His Tyr 180 185 190 Lys Lys Pro Gly Tyr Asn Gly Ser Cys Phe Asn Val Glu Ile Lys Arg 195 200 205 Asn Asp Asp Leu Ser Trp Leu Trp Asn Glu Ser Thr Ala Leu Tyr Pro 210 215 220 Ser Ile Tyr Leu Asn Thr Gln Gln Ser Pro Val Ala Ala Thr Leu Tyr 225 230 235 240 Val Arg Asn Arg Val Arg Glu Ala Ile Arg Val Ser Lys Ile Pro Asp 245 250 255 Ala Lys Ser Pro Leu Pro Val Phe Ala Tyr Thr Arg Ile Val Phe Thr 260 265 270 Asp Gln Val Leu Lys Phe Leu Ser Gln Asp Glu Leu Val Tyr Thr Phe 275 280 285 Gly Glu Thr Val Ala Leu Gly Ala Ser Gly Ile Val Ile Trp Gly Thr 290 295 300 Trp Glu Asn Thr Arg Thr Lys Glu Ser Cys Gln Ala Ile Lys Glu Tyr 305 310 315 320 Met Asp Thr Thr Leu Asn Pro Tyr Ile Ile Asn Val Thr Leu Ala Ala 325 330 335 Lys Met Cys Ser Gln Val Leu Cys Gln Glu Gln Gly Val Cys Ile Arg 340 345 350 Lys Asn Trp Asn Ser Ser Asp Tyr Leu His Leu Asn Pro Asp Asn Phe 355 360 365 Ala Ile Gln Leu Glu Lys Gly Gly Lys Phe Thr Val Arg Gly Lys Pro 370 375 380 Thr Leu Glu Asp Leu Glu Gln Phe Ser Glu Lys Phe Tyr Cys Ser Cys 385 390 395 400 Tyr Ser Thr Leu Ser Cys Lys Glu Lys Ala Asp Val Lys Asp Thr Asp 405 410 415 Ala Val Asp Val Cys Ile Ala Asp Gly Val Cys Ile Asp Ala Phe Leu 420 425 430 Lys Pro Pro Met Glu Thr Glu 435 ＜210＞ 38 <211> 454 ＜212＞ PRT ＜213＞ Artificial sequence <220> ＜223＞ Synthetic sequence ＜400＞ 38 Asn Phe Arg Ala Pro Pro Val Ile Pro Asn Val Pro Phe Leu Trp Ala 1 5 10 15 Trp Asn Ala Pro Ser Glu Phe Cys Leu Gly Lys Phe Asp Glu Pro Leu 20 25 30 Asp Met Ser Leu Phe Ser Phe Ile Gly Ser Pro Arg Ile Asn Ala Thr 35 40 45 Gly Gln Gly Val Thr Ile Phe Tyr Val Asp Arg Leu Gly Tyr Tyr Pro 50 55 60 Tyr Ile Asp Ser Ile Thr Gly Val Thr Val Asn Gly Gly Ile Pro Gln 65 70 75 80 Lys Ile Ser Leu Gln Asp His Leu Asp Lys Ala Lys Lys Asp Ile Thr 85 90 95 Phe Tyr Met Pro Val Asp Asn Leu Gly Met Ala Val Ile Asp Trp Glu 100 105 110 Glu Trp Arg Pro Thr Trp Ala Arg Asn Trp Lys Pro Lys Asp Val Tyr 115 120 125 Lys Asn Arg Ser Ile Glu Leu Val Gln Gln Gln Asn Val Gln Leu Ser 130 135 140 Leu Thr Glu Ala Thr Glu Lys Ala Lys Gln Glu Phe Glu Lys Ala Gly 145 150 155 160 Lys Asp Phe Leu Val Glu Thr Ile Lys Leu Gly Lys Leu Leu Arg Pro 165 170 175 Asn His Leu Trp Gly Tyr Tyr Leu Phe Pro Asp Cys Tyr Asn His His 180 185 190 Tyr Lys Lys Pro Gly Tyr Asn Gly Ser Cys Phe Asn Val Glu Ile Lys 195 200 205 Arg Asn Asp Asp Leu Ser Trp Leu Trp Asn Glu Ser Thr Ala Leu Tyr 210 215 220 Pro Ser Ile Tyr Leu Asn Thr Gln Gln Ser Pro Val Ala Ala Thr Leu 225 230 235 240 Tyr Val Arg Asn Arg Val Arg Glu Ala Ile Arg Val Ser Lys Ile Pro 245 250 255 Asp Ala Lys Ser Pro Leu Pro Val Phe Ala Tyr Thr Arg Ile Val Phe 260 265 270 Thr Asp Gln Val Leu Lys Phe Leu Ser Gln Asp Glu Leu Val Tyr Thr 275 280 285 Phe Gly Glu Thr Val Ala Leu Gly Ala Ser Gly Ile Val Ile Trp Gly 290 295 300 Thr Leu Ser Ile Thr Arg Thr Lys Glu Ser Cys Gln Ala Ile Lys Glu 305 310 315 320 Tyr Met Asp Thr Thr Leu Asn Pro Tyr Ile Ile Asn Val Thr Leu Ala 325 330 335 Ala Lys Met Cys Ser Gln Val Leu Cys Gln Glu Gln Gly Val Cys Ile 340 345 350 Arg Lys Asn Trp Asn Ser Ser Asp Tyr Leu His Leu Asn Pro Asp Asn 355 360 365 Phe Ala Ile Gln Leu Glu Lys Gly Gly Lys Phe Thr Val Arg Gly Lys 370 375 380 Pro Thr Leu Glu Asp Leu Glu Gln Phe Ser Glu Lys Phe Tyr Cys Ser 385 390 395 400 Cys Tyr Ser Thr Leu Ser Cys Lys Glu Lys Ala Asp Val Lys Asp Thr 405 410 415 Asp Ala Val Asp Val Cys Ile Ala Asp Gly Val Cys Ile Asp Ala Phe 420 425 430 Leu Lys Pro Pro Met Glu Thr Glu Glu Pro Gln Ile Phe Tyr Asn Ala 435 440 445 Ser Pro Ser Thr Leu Ser 450 ＜210＞ 39 ＜211＞ 452 ＜212＞ PRT ＜213＞ Artificial sequence <220> ＜223＞ Synthetic sequence ＜400＞ 39 Arg Ala Pro Pro Val Ile Pro Asn Val Pro Phe Leu Trp Ala Trp Asn 1 5 10 15 Ala Pro Ser Glu Phe Cys Leu Gly Lys Phe Asp Glu Pro Leu Asp Met 20 25 30 Ser Leu Phe Ser Phe Ile Gly Ser Pro Arg Ile Asn Ala Thr Gly Gln 35 40 45 Gly Val Thr Ile Phe Tyr Val Asp Arg Leu Gly Tyr Tyr Pro Tyr Ile 50 55 60 Asp Ser Ile Thr Gly Val Thr Val Asn Gly Gly Ile Pro Gln Lys Ile 65 70 75 80 Ser Leu Gln Asp His Leu Asp Lys Ala Lys Lys Asp Ile Thr Phe Tyr 85 90 95 Met Pro Val Asp Asn Leu Gly Met Ala Val Ile Asp Trp Glu Glu Trp 100 105 110 Arg Pro Thr Trp Ala Arg Asn Trp Lys Pro Lys Asp Val Tyr Lys Asn 115 120 125 Arg Ser Ile Glu Leu Val Gln Gln Gln Asn Val Gln Leu Ser Leu Thr 130 135 140 Glu Ala Thr Glu Lys Ala Lys Gln Glu Phe Glu Lys Ala Gly Lys Asp 145 150 155 160 Phe Leu Val Glu Thr Ile Lys Leu Gly Lys Leu Leu Arg Pro Asn His 165 170 175 Leu Trp Gly Tyr Tyr Leu Phe Pro Asp Cys Tyr Asn His His Tyr Lys 180 185 190 Lys Pro Gly Tyr Asn Gly Ser Cys Phe Asn Val Glu Ile Lys Arg Asn 195 200 205 Asp Asp Leu Ser Trp Leu Trp Asn Glu Ser Thr Ala Leu Tyr Pro Ser 210 215 220 Ile Tyr Leu Asn Thr Gln Gln Ser Pro Val Ala Ala Thr Leu Tyr Val 225 230 235 240 Arg Asn Arg Val Arg Glu Ala Ile Arg Val Ser Lys Ile Pro Asp Ala 245 250 255 Lys Ser Pro Leu Pro Val Phe Ala Tyr Thr Arg Ile Val Phe Thr Asp 260 265 270 Gln Val Leu Lys Phe Leu Ser Gln Asp Glu Leu Val Tyr Thr Phe Gly 275 280 285 Glu Thr Val Ala Leu Gly Ala Ser Gly Ile Val Ile Trp Gly Thr Leu 290 295 300 Ser Ile Thr Arg Thr Lys Glu Ser Cys Gln Ala Ile Lys Glu Tyr Met 305 310 315 320 Asp Thr Thr Leu Asn Pro Tyr Ile Ile Asn Val Thr Leu Ala Ala Lys 325 330 335 Met Cys Ser Gln Val Leu Cys Gln Glu Gln Gly Val Cys Ile Arg Lys 340 345 350 Asn Trp Asn Ser Ser Asp Tyr Leu His Leu Asn Pro Asp Asn Phe Ala 355 360 365 Ile Gln Leu Glu Lys Gly Gly Lys Phe Thr Val Arg Gly Lys Pro Thr 370 375 380 Leu Glu Asp Leu Glu Gln Phe Ser Glu Lys Phe Tyr Cys Ser Cys Tyr 385 390 395 400 Ser Thr Leu Ser Cys Lys Glu Lys Ala Asp Val Lys Asp Thr Asp Ala 405 410 415 Val Asp Val Cys Ile Ala Asp Gly Val Cys Ile Asp Ala Phe Leu Lys 420 425 430 Pro Pro Met Glu Thr Glu Glu Pro Gln Ile Phe Tyr Asn Ala Ser Pro 435 440 445 Ser Thr Leu Ser 450 ＜210＞ 40 ＜211＞ 450 ＜212＞ PRT ＜213＞ Artificial sequence <220> ＜223＞ Synthetic sequence ＜400＞ 40 Pro Pro Val Ile Pro Asn Val Pro Phe Leu Trp Ala Trp Asn Ala Pro 1 5 10 15 Ser Glu Phe Cys Leu Gly Lys Phe Asp Glu Pro Leu Asp Met Ser Leu 20 25 30 Phe Ser Phe Ile Gly Ser Pro Arg Ile Asn Ala Thr Gly Gln Gly Val 35 40 45 Thr Ile Phe Tyr Val Asp Arg Leu Gly Tyr Tyr Pro Tyr Ile Asp Ser 50 55 60 Ile Thr Gly Val Thr Val Asn Gly Gly Ile Pro Gln Lys Ile Ser Leu 65 70 75 80 Gln Asp His Leu Asp Lys Ala Lys Lys Asp Ile Thr Phe Tyr Met Pro 85 90 95 Val Asp Asn Leu Gly Met Ala Val Ile Asp Trp Glu Glu Trp Arg Pro 100 105 110 Thr Trp Ala Arg Asn Trp Lys Pro Lys Asp Val Tyr Lys Asn Arg Ser 115 120 125 Ile Glu Leu Val Gln Gln Gln Asn Val Gln Leu Ser Leu Thr Glu Ala 130 135 140 Thr Glu Lys Ala Lys Gln Glu Phe Glu Lys Ala Gly Lys Asp Phe Leu 145 150 155 160 Val Glu Thr Ile Lys Leu Gly Lys Leu Leu Arg Pro Asn His Leu Trp 165 170 175 Gly Tyr Tyr Leu Phe Pro Asp Cys Tyr Asn His His Tyr Lys Lys Pro 180 185 190 Gly Tyr Asn Gly Ser Cys Phe Asn Val Glu Ile Lys Arg Asn Asp Asp 195 200 205 Leu Ser Trp Leu Trp Asn Glu Ser Thr Ala Leu Tyr Pro Ser Ile Tyr 210 215 220 Leu Asn Thr Gln Gln Ser Pro Val Ala Ala Thr Leu Tyr Val Arg Asn 225 230 235 240 Arg Val Arg Glu Ala Ile Arg Val Ser Lys Ile Pro Asp Ala Lys Ser 245 250 255 Pro Leu Pro Val Phe Ala Tyr Thr Arg Ile Val Phe Thr Asp Gln Val 260 265 270 Leu Lys Phe Leu Ser Gln Asp Glu Leu Val Tyr Thr Phe Gly Glu Thr 275 280 285 Val Ala Leu Gly Ala Ser Gly Ile Val Ile Trp Gly Thr Leu Ser Ile 290 295 300 Thr Arg Thr Lys Glu Ser Cys Gln Ala Ile Lys Glu Tyr Met Asp Thr 305 310 315 320 Thr Leu Asn Pro Tyr Ile Ile Asn Val Thr Leu Ala Ala Lys Met Cys 325 330 335 Ser Gln Val Leu Cys Gln Glu Gln Gly Val Cys Ile Arg Lys Asn Trp 340 345 350 Asn Ser Ser Asp Tyr Leu His Leu Asn Pro Asp Asn Phe Ala Ile Gln 355 360 365 Leu Glu Lys Gly Gly Lys Phe Thr Val Arg Gly Lys Pro Thr Leu Glu 370 375 380 Asp Leu Glu Gln Phe Ser Glu Lys Phe Tyr Cys Ser Cys Tyr Ser Thr 385 390 395 400 Leu Ser Cys Lys Glu Lys Ala Asp Val Lys Asp Thr Asp Ala Val Asp 405 410 415 Val Cys Ile Ala Asp Gly Val Cys Ile Asp Ala Phe Leu Lys Pro Pro 420 425 430 Met Glu Thr Glu Glu Pro Gln Ile Phe Tyr Asn Ala Ser Pro Ser Thr 435 440 445 Leu Ser 450 ＜210＞ 41 ＜211＞ 428 ＜212＞ PRT ＜213＞ Artificial sequence <220> ＜223＞ Synthetic sequence ＜400＞ 41 Phe Arg Ala Pro Pro Val Ile Pro Asn Val Pro Phe Leu Trp Ala Trp 1 5 10 15 Asn Ala Pro Ser Glu Phe Cys Leu Gly Lys Phe Asp Glu Pro Leu Asp 20 25 30 Met Ser Leu Phe Ser Phe Ile Gly Ser Pro Arg Ile Asn Ala Thr Gly 35 40 45 Gln Gly Val Thr Ile Phe Tyr Val Asp Arg Leu Gly Tyr Tyr Pro Tyr 50 55 60 Ile Asp Ser Ile Thr Gly Val Thr Val Asn Gly Gly Ile Pro Gln Lys 65 70 75 80 Ile Ser Leu Gln Asp His Leu Asp Lys Ala Lys Lys Asp Ile Thr Phe 85 90 95 Tyr Met Pro Val Asp Asn Leu Gly Met Ala Val Ile Asp Trp Glu Glu 100 105 110 Trp Arg Pro Thr Trp Ala Arg Asn Trp Lys Pro Lys Asp Val Tyr Lys 115 120 125 Asn Arg Ser Ile Glu Leu Val Gln Gln Gln Asn Val Gln Leu Ser Leu 130 135 140 Thr Glu Ala Thr Glu Lys Ala Lys Gln Glu Phe Glu Lys Ala Gly Lys 145 150 155 160 Asp Phe Leu Val Glu Thr Ile Lys Leu Gly Lys Leu Leu Arg Pro Asn 165 170 175 His Leu Trp Gly Tyr Tyr Leu Phe Pro Asp Cys Tyr Asn His His Tyr 180 185 190 Lys Lys Pro Gly Tyr Asn Gly Ser Cys Phe Asn Val Glu Ile Lys Arg 195 200 205 Asn Asp Asp Leu Ser Trp Leu Trp Asn Glu Ser Thr Ala Leu Tyr Pro 210 215 220 Ser Ile Tyr Leu Asn Thr Gln Gln Ser Pro Val Ala Ala Thr Leu Tyr 225 230 235 240 Val Arg Asn Arg Val Arg Glu Ala Ile Arg Val Ser Lys Ile Pro Asp 245 250 255 Ala Lys Ser Pro Leu Pro Val Phe Ala Tyr Thr Arg Ile Val Phe Thr 260 265 270 Asp Gln Val Leu Lys Phe Leu Ser Gln Asp Glu Leu Val Tyr Thr Phe 275 280 285 Gly Glu Thr Val Ala Leu Gly Ala Ser Gly Ile Val Ile Trp Gly Ser 290 295 300 Trp Glu Asn Thr Arg Thr Lys Glu Ser Cys Gln Ala Ile Lys Glu Tyr 305 310 315 320 Met Asp Thr Thr Leu Asn Pro Tyr Ile Ile Asn Val Thr Leu Ala Ala 325 330 335 Lys Met Cys Ser Gln Val Leu Cys Gln Glu Gln Gly Val Cys Ile Arg 340 345 350 Lys Asn Trp Asn Ser Ser Asp Tyr Leu His Leu Asn Pro Asp Asn Phe 355 360 365 Ala Ile Gln Leu Glu Lys Gly Gly Lys Phe Thr Val Arg Gly Lys Pro 370 375 380 Thr Leu Glu Asp Leu Glu Gln Phe Ser Glu Lys Phe Tyr Cys Ser Cys 385 390 395 400 Tyr Ser Thr Leu Ser Cys Lys Glu Lys Ala Asp Val Lys Asp Thr Asp 405 410 415 Ala Val Asp Val Cys Ile Ala Asp Gly Val Cys Ile 420 425 ＜210＞ 42 ＜211＞ 429 ＜212＞ PRT ＜213＞ Artificial sequence <220> ＜223＞ Synthetic sequence ＜400＞ 42 Phe Arg Ala Pro Pro Val Ile Pro Asn Val Pro Phe Leu Trp Ala Trp 1 5 10 15 Asn Ala Pro Ser Glu Phe Cys Leu Gly Lys Phe Asp Glu Pro Leu Asp 20 25 30 Met Ser Leu Phe Ser Phe Ile Gly Ser Pro Arg Ile Asn Ala Thr Gly 35 40 45 Gln Gly Val Thr Ile Phe Tyr Val Asp Arg Leu Gly Tyr Tyr Pro Tyr 50 55 60 Ile Asp Ser Ile Thr Gly Val Thr Val Asn Gly Gly Ile Pro Gln Lys 65 70 75 80 Ile Ser Leu Gln Asp His Leu Asp Lys Ala Lys Lys Asp Ile Thr Phe 85 90 95 Tyr Met Pro Val Asp Asn Leu Gly Met Ala Val Ile Asp Trp Glu Glu 100 105 110 Trp Arg Pro Thr Trp Ala Arg Asn Trp Lys Pro Lys Asp Val Tyr Lys 115 120 125 Asn Arg Ser Ile Glu Leu Val Gln Gln Gln Asn Val Gln Leu Ser Leu 130 135 140 Thr Glu Ala Thr Glu Lys Ala Lys Gln Glu Phe Glu Lys Ala Gly Lys 145 150 155 160 Asp Phe Leu Val Glu Thr Ile Lys Leu Gly Lys Leu Leu Arg Pro Asn 165 170 175 His Leu Trp Gly Tyr Tyr Leu Phe Pro Asp Cys Tyr Asn His His Tyr 180 185 190 Lys Lys Pro Gly Tyr Asn Gly Ser Cys Phe Asn Val Glu Ile Lys Arg 195 200 205 Asn Asp Asp Leu Ser Trp Leu Trp Asn Glu Ser Thr Ala Leu Tyr Pro 210 215 220 Ser Ile Tyr Leu Asn Thr Gln Gln Ser Pro Val Ala Ala Thr Leu Tyr 225 230 235 240 Val Arg Asn Arg Val Arg Glu Ala Ile Arg Val Ser Lys Ile Pro Asp 245 250 255 Ala Lys Ser Pro Leu Pro Val Phe Ala Tyr Thr Arg Ile Val Phe Thr 260 265 270 Asp Gln Val Leu Lys Phe Leu Ser Gln Asp Glu Leu Val Tyr Thr Phe 275 280 285 Gly Glu Thr Val Ala Leu Gly Ala Ser Gly Ile Val Ile Trp Gly Ser 290 295 300 Trp Glu Asn Thr Arg Thr Lys Glu Ser Cys Gln Ala Ile Lys Glu Tyr 305 310 315 320 Met Asp Thr Thr Leu Asn Pro Tyr Ile Ile Asn Val Thr Leu Ala Ala 325 330 335 Lys Met Cys Ser Gln Val Leu Cys Gln Glu Gln Gly Val Cys Ile Arg 340 345 350 Lys Asn Trp Asn Ser Ser Asp Tyr Leu His Leu Asn Pro Asp Asn Phe 355 360 365 Ala Ile Gln Leu Glu Lys Gly Gly Lys Phe Thr Val Arg Gly Lys Pro 370 375 380 Thr Leu Glu Asp Leu Glu Gln Phe Ser Glu Lys Phe Tyr Cys Ser Cys 385 390 395 400 Tyr Ser Thr Leu Ser Cys Lys Glu Lys Ala Asp Val Lys Asp Thr Asp 405 410 415 Ala Val Asp Val Cys Ile Ala Asp Gly Val Cys Ile Asp 420 425 ＜210＞ 43 ＜211＞ 430 ＜212＞ PRT ＜213＞ Artificial sequence <220> ＜223＞ Synthetic sequence ＜400＞ 43 Phe Arg Ala Pro Pro Val Ile Pro Asn Val Pro Phe Leu Trp Ala Trp 1 5 10 15 Asn Ala Pro Ser Glu Phe Cys Leu Gly Lys Phe Asp Glu Pro Leu Asp 20 25 30 Met Ser Leu Phe Ser Phe Ile Gly Ser Pro Arg Ile Asn Ala Thr Gly 35 40 45 Gln Gly Val Thr Ile Phe Tyr Val Asp Arg Leu Gly Tyr Tyr Pro Tyr 50 55 60 Ile Asp Ser Ile Thr Gly Val Thr Val Asn Gly Gly Ile Pro Gln Lys 65 70 75 80 Ile Ser Leu Gln Asp His Leu Asp Lys Ala Lys Lys Asp Ile Thr Phe 85 90 95 Tyr Met Pro Val Asp Asn Leu Gly Met Ala Val Ile Asp Trp Glu Glu 100 105 110 Trp Arg Pro Thr Trp Ala Arg Asn Trp Lys Pro Lys Asp Val Tyr Lys 115 120 125 Asn Arg Ser Ile Glu Leu Val Gln Gln Gln Asn Val Gln Leu Ser Leu 130 135 140 Thr Glu Ala Thr Glu Lys Ala Lys Gln Glu Phe Glu Lys Ala Gly Lys 145 150 155 160 Asp Phe Leu Val Glu Thr Ile Lys Leu Gly Lys Leu Leu Arg Pro Asn 165 170 175 His Leu Trp Gly Tyr Tyr Leu Phe Pro Asp Cys Tyr Asn His His Tyr 180 185 190 Lys Lys Pro Gly Tyr Asn Gly Ser Cys Phe Asn Val Glu Ile Lys Arg 195 200 205 Asn Asp Asp Leu Ser Trp Leu Trp Asn Glu Ser Thr Ala Leu Tyr Pro 210 215 220 Ser Ile Tyr Leu Asn Thr Gln Gln Ser Pro Val Ala Ala Thr Leu Tyr 225 230 235 240 Val Arg Asn Arg Val Arg Glu Ala Ile Arg Val Ser Lys Ile Pro Asp 245 250 255 Ala Lys Ser Pro Leu Pro Val Phe Ala Tyr Thr Arg Ile Val Phe Thr 260 265 270 Asp Gln Val Leu Lys Phe Leu Ser Gln Asp Glu Leu Val Tyr Thr Phe 275 280 285 Gly Glu Thr Val Ala Leu Gly Ala Ser Gly Ile Val Ile Trp Gly Ser 290 295 300 Trp Glu Asn Thr Arg Thr Lys Glu Ser Cys Gln Ala Ile Lys Glu Tyr 305 310 315 320 Met Asp Thr Thr Leu Asn Pro Tyr Ile Ile Asn Val Thr Leu Ala Ala 325 330 335 Lys Met Cys Ser Gln Val Leu Cys Gln Glu Gln Gly Val Cys Ile Arg 340 345 350 Lys Asn Trp Asn Ser Ser Asp Tyr Leu His Leu Asn Pro Asp Asn Phe 355 360 365 Ala Ile Gln Leu Glu Lys Gly Gly Lys Phe Thr Val Arg Gly Lys Pro 370 375 380 Thr Leu Glu Asp Leu Glu Gln Phe Ser Glu Lys Phe Tyr Cys Ser Cys 385 390 395 400 Tyr Ser Thr Leu Ser Cys Lys Glu Lys Ala Asp Val Lys Asp Thr Asp 405 410 415 Ala Val Asp Val Cys Ile Ala Asp Gly Val Cys Ile Asp Ala 420 425 430 ＜210＞ 44 ＜211＞ 431 ＜212＞ PRT ＜213＞ Artificial sequence <220> ＜223＞ Synthetic sequence ＜400＞ 44 Phe Arg Ala Pro Pro Val Ile Pro Asn Val Pro Phe Leu Trp Ala Trp 1 5 10 15 Asn Ala Pro Ser Glu Phe Cys Leu Gly Lys Phe Asp Glu Pro Leu Asp 20 25 30 Met Ser Leu Phe Ser Phe Ile Gly Ser Pro Arg Ile Asn Ala Thr Gly 35 40 45 Gln Gly Val Thr Ile Phe Tyr Val Asp Arg Leu Gly Tyr Tyr Pro Tyr 50 55 60 Ile Asp Ser Ile Thr Gly Val Thr Val Asn Gly Gly Ile Pro Gln Lys 65 70 75 80 Ile Ser Leu Gln Asp His Leu Asp Lys Ala Lys Lys Asp Ile Thr Phe 85 90 95 Tyr Met Pro Val Asp Asn Leu Gly Met Ala Val Ile Asp Trp Glu Glu 100 105 110 Trp Arg Pro Thr Trp Ala Arg Asn Trp Lys Pro Lys Asp Val Tyr Lys 115 120 125 Asn Arg Ser Ile Glu Leu Val Gln Gln Gln Asn Val Gln Leu Ser Leu 130 135 140 Thr Glu Ala Thr Glu Lys Ala Lys Gln Glu Phe Glu Lys Ala Gly Lys 145 150 155 160 Asp Phe Leu Val Glu Thr Ile Lys Leu Gly Lys Leu Leu Arg Pro Asn 165 170 175 His Leu Trp Gly Tyr Tyr Leu Phe Pro Asp Cys Tyr Asn His His Tyr 180 185 190 Lys Lys Pro Gly Tyr Asn Gly Ser Cys Phe Asn Val Glu Ile Lys Arg 195 200 205 Asn Asp Asp Leu Ser Trp Leu Trp Asn Glu Ser Thr Ala Leu Tyr Pro 210 215 220 Ser Ile Tyr Leu Asn Thr Gln Gln Ser Pro Val Ala Ala Thr Leu Tyr 225 230 235 240 Val Arg Asn Arg Val Arg Glu Ala Ile Arg Val Ser Lys Ile Pro Asp 245 250 255 Ala Lys Ser Pro Leu Pro Val Phe Ala Tyr Thr Arg Ile Val Phe Thr 260 265 270 Asp Gln Val Leu Lys Phe Leu Ser Gln Asp Glu Leu Val Tyr Thr Phe 275 280 285 Gly Glu Thr Val Ala Leu Gly Ala Ser Gly Ile Val Ile Trp Gly Ser 290 295 300 Trp Glu Asn Thr Arg Thr Lys Glu Ser Cys Gln Ala Ile Lys Glu Tyr 305 310 315 320 Met Asp Thr Thr Leu Asn Pro Tyr Ile Ile Asn Val Thr Leu Ala Ala 325 330 335 Lys Met Cys Ser Gln Val Leu Cys Gln Glu Gln Gly Val Cys Ile Arg 340 345 350 Lys Asn Trp Asn Ser Ser Asp Tyr Leu His Leu Asn Pro Asp Asn Phe 355 360 365 Ala Ile Gln Leu Glu Lys Gly Gly Lys Phe Thr Val Arg Gly Lys Pro 370 375 380 Thr Leu Glu Asp Leu Glu Gln Phe Ser Glu Lys Phe Tyr Cys Ser Cys 385 390 395 400 Tyr Ser Thr Leu Ser Cys Lys Glu Lys Ala Asp Val Lys Asp Thr Asp 405 410 415 Ala Val Asp Val Cys Ile Ala Asp Gly Val Cys Ile Asp Ala Phe 420 425 430 ＜210＞ 45 <211> 441 ＜212＞ PRT ＜213＞ Artificial sequence <220> ＜223＞ Synthetic sequence ＜400＞ 45 Phe Arg Ala Pro Pro Val Ile Pro Asn Val Pro Phe Leu Trp Ala Trp 1 5 10 15 Asn Ala Pro Ser Glu Phe Cys Leu Gly Lys Phe Asp Glu Pro Leu Asp 20 25 30 Met Ser Leu Phe Ser Phe Ile Gly Ser Pro Arg Ile Asn Ala Thr Gly 35 40 45 Gln Gly Val Thr Ile Phe Tyr Val Asp Arg Leu Gly Tyr Tyr Pro Tyr 50 55 60 Ile Asp Ser Ile Thr Gly Val Thr Val Asn Gly Gly Ile Pro Gln Lys 65 70 75 80 Ile Ser Leu Gln Asp His Leu Asp Lys Ala Lys Lys Asp Ile Thr Phe 85 90 95 Tyr Met Pro Val Asp Asn Leu Gly Met Ala Val Ile Asp Trp Glu Glu 100 105 110 Trp Arg Pro Thr Trp Ala Arg Asn Trp Lys Pro Lys Asp Val Tyr Lys 115 120 125 Asn Arg Ser Ile Glu Leu Val Gln Gln Gln Asn Val Gln Leu Ser Leu 130 135 140 Thr Glu Ala Thr Glu Lys Ala Lys Gln Glu Phe Glu Lys Ala Gly Lys 145 150 155 160 Asp Phe Leu Val Glu Thr Ile Lys Leu Gly Lys Leu Leu Arg Pro Asn 165 170 175 His Leu Trp Gly Tyr Tyr Leu Phe Pro Asp Cys Tyr Asn His His Tyr 180 185 190 Lys Lys Pro Gly Tyr Asn Gly Ser Cys Phe Asn Val Glu Ile Lys Arg 195 200 205 Asn Asp Asp Leu Ser Trp Leu Trp Asn Glu Ser Thr Ala Leu Tyr Pro 210 215 220 Ser Ile Tyr Leu Asn Thr Gln Gln Ser Pro Val Ala Ala Thr Leu Tyr 225 230 235 240 Val Arg Asn Arg Val Arg Glu Ala Ile Arg Val Ser Lys Ile Pro Asp 245 250 255 Ala Lys Ser Pro Leu Pro Val Phe Ala Tyr Thr Arg Ile Val Phe Thr 260 265 270 Asp Gln Val Leu Lys Phe Leu Ser Gln Asp Glu Leu Val Tyr Thr Phe 275 280 285 Gly Glu Thr Val Ala Leu Gly Ala Ser Gly Ile Val Ile Trp Gly Thr 290 295 300 Trp Glu Asn Thr Arg Thr Lys Glu Ser Cys Gln Ala Ile Lys Glu Tyr 305 310 315 320 Met Asp Thr Thr Leu Asn Pro Tyr Ile Ile Asn Val Thr Leu Ala Ala 325 330 335 Lys Met Cys Ser Gln Val Leu Cys Gln Glu Gln Gly Val Cys Ile Arg 340 345 350 Lys Asn Trp Asn Ser Ser Asp Tyr Leu His Leu Asn Pro Asp Asn Phe 355 360 365 Ala Ile Gln Leu Glu Lys Gly Gly Lys Phe Thr Val Arg Gly Lys Pro 370 375 380 Thr Leu Glu Asp Leu Glu Gln Phe Ser Glu Lys Phe Tyr Cys Ser Cys 385 390 395 400 Tyr Ser Thr Leu Ser Cys Lys Glu Lys Ala Asp Val Lys Asp Thr Asp 405 410 415 Ala Val Asp Val Cys Ile Ala Asp Gly Val Cys Ile Asp Ala Phe Leu 420 425 430 Lys Pro Pro Met Glu Thr Glu Glu Pro 435 440 ＜210＞ 46 <211> 443 ＜212＞ PRT ＜213＞ Artificial sequence <220> ＜223＞ Synthetic sequence ＜400＞ 46 Phe Arg Ala Pro Pro Val Ile Pro Asn Val Pro Phe Leu Trp Ala Trp 1 5 10 15 Asn Ala Pro Ser Glu Phe Cys Leu Gly Lys Phe Asp Glu Pro Leu Asp 20 25 30 Met Ser Leu Phe Ser Phe Ile Gly Ser Pro Arg Ile Asn Ala Thr Gly 35 40 45 Gln Gly Val Thr Ile Phe Tyr Val Asp Arg Leu Gly Tyr Tyr Pro Tyr 50 55 60 Ile Asp Ser Ile Thr Gly Val Thr Val Asn Gly Gly Ile Pro Gln Lys 65 70 75 80 Ile Ser Leu Gln Asp His Leu Asp Lys Ala Lys Lys Asp Ile Thr Phe 85 90 95 Tyr Met Pro Val Asp Asn Leu Gly Met Ala Val Ile Asp Trp Glu Glu 100 105 110 Trp Arg Pro Thr Trp Ala Arg Asn Trp Lys Pro Lys Asp Val Tyr Lys 115 120 125 Asn Arg Ser Ile Glu Leu Val Gln Gln Gln Asn Val Gln Leu Ser Leu 130 135 140 Thr Glu Ala Thr Glu Lys Ala Lys Gln Glu Phe Glu Lys Ala Gly Lys 145 150 155 160 Asp Phe Leu Val Glu Thr Ile Lys Leu Gly Lys Leu Leu Arg Pro Asn 165 170 175 His Leu Trp Gly Tyr Tyr Leu Phe Pro Asp Cys Tyr Asn His His Tyr 180 185 190 Lys Lys Pro Gly Tyr Asn Gly Ser Cys Phe Asn Val Glu Ile Lys Arg 195 200 205 Asn Asp Asp Leu Ser Trp Leu Trp Asn Glu Ser Thr Ala Leu Tyr Pro 210 215 220 Ser Ile Tyr Leu Asn Thr Gln Gln Ser Pro Val Ala Ala Thr Leu Tyr 225 230 235 240 Val Arg Asn Arg Val Arg Glu Ala Ile Arg Val Ser Lys Ile Pro Asp 245 250 255 Ala Lys Ser Pro Leu Pro Val Phe Ala Tyr Thr Arg Ile Val Phe Thr 260 265 270 Asp Gln Val Leu Lys Phe Leu Ser Gln Asp Glu Leu Val Tyr Thr Phe 275 280 285 Gly Glu Thr Val Ala Leu Gly Ala Ser Gly Ile Val Ile Trp Gly Thr 290 295 300 Trp Glu Asn Thr Arg Thr Lys Glu Ser Cys Gln Ala Ile Lys Glu Tyr 305 310 315 320 Met Asp Thr Thr Leu Asn Pro Tyr Ile Ile Asn Val Thr Leu Ala Ala 325 330 335 Lys Met Cys Ser Gln Val Leu Cys Gln Glu Gln Gly Val Cys Ile Arg 340 345 350 Lys Asn Trp Asn Ser Ser Asp Tyr Leu His Leu Asn Pro Asp Asn Phe 355 360 365 Ala Ile Gln Leu Glu Lys Gly Gly Lys Phe Thr Val Arg Gly Lys Pro 370 375 380 Thr Leu Glu Asp Leu Glu Gln Phe Ser Glu Lys Phe Tyr Cys Ser Cys 385 390 395 400 Tyr Ser Thr Leu Ser Cys Lys Glu Lys Ala Asp Val Lys Asp Thr Asp 405 410 415 Ala Val Asp Val Cys Ile Ala Asp Gly Val Cys Ile Asp Ala Phe Leu 420 425 430 Lys Pro Pro Met Glu Thr Glu Glu Pro Gln Ile 435 440 ＜210＞ 47 ＜211＞ 445 ＜212＞ PRT ＜213＞ Artificial sequence <220> ＜223＞ Synthetic sequence ＜400＞ 47 Phe Arg Ala Pro Pro Val Ile Pro Asn Val Pro Phe Leu Trp Ala Trp 1 5 10 15 Asn Ala Pro Ser Glu Phe Cys Leu Gly Lys Phe Asp Glu Pro Leu Asp 20 25 30 Met Ser Leu Phe Ser Phe Ile Gly Ser Pro Arg Ile Asn Ala Thr Gly 35 40 45 Gln Gly Val Thr Ile Phe Tyr Val Asp Arg Leu Gly Tyr Tyr Pro Tyr 50 55 60 Ile Asp Ser Ile Thr Gly Val Thr Val Asn Gly Gly Ile Pro Gln Lys 65 70 75 80 Ile Ser Leu Gln Asp His Leu Asp Lys Ala Lys Lys Asp Ile Thr Phe 85 90 95 Tyr Met Pro Val Asp Asn Leu Gly Met Ala Val Ile Asp Trp Glu Glu 100 105 110 Trp Arg Pro Thr Trp Ala Arg Asn Trp Lys Pro Lys Asp Val Tyr Lys 115 120 125 Asn Arg Ser Ile Glu Leu Val Gln Gln Gln Asn Val Gln Leu Ser Leu 130 135 140 Thr Glu Ala Thr Glu Lys Ala Lys Gln Glu Phe Glu Lys Ala Gly Lys 145 150 155 160 Asp Phe Leu Val Glu Thr Ile Lys Leu Gly Lys Leu Leu Arg Pro Asn 165 170 175 His Leu Trp Gly Tyr Tyr Leu Phe Pro Asp Cys Tyr Asn His His Tyr 180 185 190 Lys Lys Pro Gly Tyr Asn Gly Ser Cys Phe Asn Val Glu Ile Lys Arg 195 200 205 Asn Asp Asp Leu Ser Trp Leu Trp Asn Glu Ser Thr Ala Leu Tyr Pro 210 215 220 Ser Ile Tyr Leu Asn Thr Gln Gln Ser Pro Val Ala Ala Thr Leu Tyr 225 230 235 240 Val Arg Asn Arg Val Arg Glu Ala Ile Arg Val Ser Lys Ile Pro Asp 245 250 255 Ala Lys Ser Pro Leu Pro Val Phe Ala Tyr Thr Arg Ile Val Phe Thr 260 265 270 Asp Gln Val Leu Lys Phe Leu Ser Gln Asp Glu Leu Val Tyr Thr Phe 275 280 285 Gly Glu Thr Val Ala Leu Gly Ala Ser Gly Ile Val Ile Trp Gly Thr 290 295 300 Trp Glu Asn Thr Arg Thr Lys Glu Ser Cys Gln Ala Ile Lys Glu Tyr 305 310 315 320 Met Asp Thr Thr Leu Asn Pro Tyr Ile Ile Asn Val Thr Leu Ala Ala 325 330 335 Lys Met Cys Ser Gln Val Leu Cys Gln Glu Gln Gly Val Cys Ile Arg 340 345 350 Lys Asn Trp Asn Ser Ser Asp Tyr Leu His Leu Asn Pro Asp Asn Phe 355 360 365 Ala Ile Gln Leu Glu Lys Gly Gly Lys Phe Thr Val Arg Gly Lys Pro 370 375 380 Thr Leu Glu Asp Leu Glu Gln Phe Ser Glu Lys Phe Tyr Cys Ser Cys 385 390 395 400 Tyr Ser Thr Leu Ser Cys Lys Glu Lys Ala Asp Val Lys Asp Thr Asp 405 410 415 Ala Val Asp Val Cys Ile Ala Asp Gly Val Cys Ile Asp Ala Phe Leu 420 425 430 Lys Pro Pro Met Glu Thr Glu Glu Pro Gln Ile Phe Tyr 435 440 445 ＜210＞ 48 ＜211＞ 447 ＜212＞ PRT ＜213＞ Artificial sequence <220> ＜223＞ Synthetic sequence ＜400＞ 48 Phe Arg Ala Pro Pro Val Ile Pro Asn Val Pro Phe Leu Trp Ala Trp 1 5 10 15 Asn Ala Pro Ser Glu Phe Cys Leu Gly Lys Phe Asp Glu Pro Leu Asp 20 25 30 Met Ser Leu Phe Ser Phe Ile Gly Ser Pro Arg Ile Asn Ala Thr Gly 35 40 45 Gln Gly Val Thr Ile Phe Tyr Val Asp Arg Leu Gly Tyr Tyr Pro Tyr 50 55 60 Ile Asp Ser Ile Thr Gly Val Thr Val Asn Gly Gly Ile Pro Gln Lys 65 70 75 80 Ile Ser Leu Gln Asp His Leu Asp Lys Ala Lys Lys Asp Ile Thr Phe 85 90 95 Tyr Met Pro Val Asp Asn Leu Gly Met Ala Val Ile Asp Trp Glu Glu 100 105 110 Trp Arg Pro Thr Trp Ala Arg Asn Trp Lys Pro Lys Asp Val Tyr Lys 115 120 125 Asn Arg Ser Ile Glu Leu Val Gln Gln Gln Asn Val Gln Leu Ser Leu 130 135 140 Thr Glu Ala Thr Glu Lys Ala Lys Gln Glu Phe Glu Lys Ala Gly Lys 145 150 155 160 Asp Phe Leu Val Glu Thr Ile Lys Leu Gly Lys Leu Leu Arg Pro Asn 165 170 175 His Leu Trp Gly Tyr Tyr Leu Phe Pro Asp Cys Tyr Asn His His Tyr 180 185 190 Lys Lys Pro Gly Tyr Asn Gly Ser Cys Phe Asn Val Glu Ile Lys Arg 195 200 205 Asn Asp Asp Leu Ser Trp Leu Trp Asn Glu Ser Thr Ala Leu Tyr Pro 210 215 220 Ser Ile Tyr Leu Asn Thr Gln Gln Ser Pro Val Ala Ala Thr Leu Tyr 225 230 235 240 Val Arg Asn Arg Val Arg Glu Ala Ile Arg Val Ser Lys Ile Pro Asp 245 250 255 Ala Lys Ser Pro Leu Pro Val Phe Ala Tyr Thr Arg Ile Val Phe Thr 260 265 270 Asp Gln Val Leu Lys Phe Leu Ser Gln Asp Glu Leu Val Tyr Thr Phe 275 280 285 Gly Glu Thr Val Ala Leu Gly Ala Ser Gly Ile Val Ile Trp Gly Thr 290 295 300 Trp Glu Asn Thr Arg Thr Lys Glu Ser Cys Gln Ala Ile Lys Glu Tyr 305 310 315 320 Met Asp Thr Thr Leu Asn Pro Tyr Ile Ile Asn Val Thr Leu Ala Ala 325 330 335 Lys Met Cys Ser Gln Val Leu Cys Gln Glu Gln Gly Val Cys Ile Arg 340 345 350 Lys Asn Trp Asn Ser Ser Asp Tyr Leu His Leu Asn Pro Asp Asn Phe 355 360 365 Ala Ile Gln Leu Glu Lys Gly Gly Lys Phe Thr Val Arg Gly Lys Pro 370 375 380 Thr Leu Glu Asp Leu Glu Gln Phe Ser Glu Lys Phe Tyr Cys Ser Cys 385 390 395 400 Tyr Ser Thr Leu Ser Cys Lys Glu Lys Ala Asp Val Lys Asp Thr Asp 405 410 415 Ala Val Asp Val Cys Ile Ala Asp Gly Val Cys Ile Asp Ala Phe Leu 420 425 430 Lys Pro Pro Met Glu Thr Glu Glu Pro Gln Ile Phe Tyr Asn Ala 435 440 445 ＜210＞ 49 ＜211＞ 449 ＜212＞ PRT ＜213＞ Artificial sequence <220> ＜223＞ Synthetic sequence ＜400＞ 49 Phe Arg Ala Pro Pro Val Ile Pro Asn Val Pro Phe Leu Trp Ala Trp 1 5 10 15 Asn Ala Pro Ser Glu Phe Cys Leu Gly Lys Phe Asp Glu Pro Leu Asp 20 25 30 Met Ser Leu Phe Ser Phe Ile Gly Ser Pro Arg Ile Asn Ala Thr Gly 35 40 45 Gln Gly Val Thr Ile Phe Tyr Val Asp Arg Leu Gly Tyr Tyr Pro Tyr 50 55 60 Ile Asp Ser Ile Thr Gly Val Thr Val Asn Gly Gly Ile Pro Gln Lys 65 70 75 80 Ile Ser Leu Gln Asp His Leu Asp Lys Ala Lys Lys Asp Ile Thr Phe 85 90 95 Tyr Met Pro Val Asp Asn Leu Gly Met Ala Val Ile Asp Trp Glu Glu 100 105 110 Trp Arg Pro Thr Trp Ala Arg Asn Trp Lys Pro Lys Asp Val Tyr Lys 115 120 125 Asn Arg Ser Ile Glu Leu Val Gln Gln Gln Asn Val Gln Leu Ser Leu 130 135 140 Thr Glu Ala Thr Glu Lys Ala Lys Gln Glu Phe Glu Lys Ala Gly Lys 145 150 155 160 Asp Phe Leu Val Glu Thr Ile Lys Leu Gly Lys Leu Leu Arg Pro Asn 165 170 175 His Leu Trp Gly Tyr Tyr Leu Phe Pro Asp Cys Tyr Asn His His Tyr 180 185 190 Lys Lys Pro Gly Tyr Asn Gly Ser Cys Phe Asn Val Glu Ile Lys Arg 195 200 205 Asn Asp Asp Leu Ser Trp Leu Trp Asn Glu Ser Thr Ala Leu Tyr Pro 210 215 220 Ser Ile Tyr Leu Asn Thr Gln Gln Ser Pro Val Ala Ala Thr Leu Tyr 225 230 235 240 Val Arg Asn Arg Val Arg Glu Ala Ile Arg Val Ser Lys Ile Pro Asp 245 250 255 Ala Lys Ser Pro Leu Pro Val Phe Ala Tyr Thr Arg Ile Val Phe Thr 260 265 270 Asp Gln Val Leu Lys Phe Leu Ser Gln Asp Glu Leu Val Tyr Thr Phe 275 280 285 Gly Glu Thr Val Ala Leu Gly Ala Ser Gly Ile Val Ile Trp Gly Thr 290 295 300 Trp Glu Asn Thr Arg Thr Lys Glu Ser Cys Gln Ala Ile Lys Glu Tyr 305 310 315 320 Met Asp Thr Thr Leu Asn Pro Tyr Ile Ile Asn Val Thr Leu Ala Ala 325 330 335 Lys Met Cys Ser Gln Val Leu Cys Gln Glu Gln Gly Val Cys Ile Arg 340 345 350 Lys Asn Trp Asn Ser Ser Asp Tyr Leu His Leu Asn Pro Asp Asn Phe 355 360 365 Ala Ile Gln Leu Glu Lys Gly Gly Lys Phe Thr Val Arg Gly Lys Pro 370 375 380 Thr Leu Glu Asp Leu Glu Gln Phe Ser Glu Lys Phe Tyr Cys Ser Cys 385 390 395 400 Tyr Ser Thr Leu Ser Cys Lys Glu Lys Ala Asp Val Lys Asp Thr Asp 405 410 415 Ala Val Asp Val Cys Ile Ala Asp Gly Val Cys Ile Asp Ala Phe Leu 420 425 430 Lys Pro Pro Met Glu Thr Glu Glu Pro Gln Ile Phe Tyr Asn Ala Ser 435 440 445 Pro ＜210＞ 50 <211> 451 ＜212＞ PRT ＜213＞ Artificial sequence <220> ＜223＞ Synthetic sequence ＜400＞ 50 Phe Arg Ala Pro Pro Val Ile Pro Asn Val Pro Phe Leu Trp Ala Trp 1 5 10 15 Asn Ala Pro Ser Glu Phe Cys Leu Gly Lys Phe Asp Glu Pro Leu Asp 20 25 30 Met Ser Leu Phe Ser Phe Ile Gly Ser Pro Arg Ile Asn Ala Thr Gly 35 40 45 Gln Gly Val Thr Ile Phe Tyr Val Asp Arg Leu Gly Tyr Tyr Pro Tyr 50 55 60 Ile Asp Ser Ile Thr Gly Val Thr Val Asn Gly Gly Ile Pro Gln Lys 65 70 75 80 Ile Ser Leu Gln Asp His Leu Asp Lys Ala Lys Lys Asp Ile Thr Phe 85 90 95 Tyr Met Pro Val Asp Asn Leu Gly Met Ala Val Ile Asp Trp Glu Glu 100 105 110 Trp Arg Pro Thr Trp Ala Arg Asn Trp Lys Pro Lys Asp Val Tyr Lys 115 120 125 Asn Arg Ser Ile Glu Leu Val Gln Gln Gln Asn Val Gln Leu Ser Leu 130 135 140 Thr Glu Ala Thr Glu Lys Ala Lys Gln Glu Phe Glu Lys Ala Gly Lys 145 150 155 160 Asp Phe Leu Val Glu Thr Ile Lys Leu Gly Lys Leu Leu Arg Pro Asn 165 170 175 His Leu Trp Gly Tyr Tyr Leu Phe Pro Asp Cys Tyr Asn His His Tyr 180 185 190 Lys Lys Pro Gly Tyr Asn Gly Ser Cys Phe Asn Val Glu Ile Lys Arg 195 200 205 Asn Asp Asp Leu Ser Trp Leu Trp Asn Glu Ser Thr Ala Leu Tyr Pro 210 215 220 Ser Ile Tyr Leu Asn Thr Gln Gln Ser Pro Val Ala Ala Thr Leu Tyr 225 230 235 240 Val Arg Asn Arg Val Arg Glu Ala Ile Arg Val Ser Lys Ile Pro Asp 245 250 255 Ala Lys Ser Pro Leu Pro Val Phe Ala Tyr Thr Arg Ile Val Phe Thr 260 265 270 Asp Gln Val Leu Lys Phe Leu Ser Gln Asp Glu Leu Val Tyr Thr Phe 275 280 285 Gly Glu Thr Val Ala Leu Gly Ala Ser Gly Ile Val Ile Trp Gly Thr 290 295 300 Trp Glu Asn Thr Arg Thr Lys Glu Ser Cys Gln Ala Ile Lys Glu Tyr 305 310 315 320 Met Asp Thr Thr Leu Asn Pro Tyr Ile Ile Asn Val Thr Leu Ala Ala 325 330 335 Lys Met Cys Ser Gln Val Leu Cys Gln Glu Gln Gly Val Cys Ile Arg 340 345 350 Lys Asn Trp Asn Ser Ser Asp Tyr Leu His Leu Asn Pro Asp Asn Phe 355 360 365 Ala Ile Gln Leu Glu Lys Gly Gly Lys Phe Thr Val Arg Gly Lys Pro 370 375 380 Thr Leu Glu Asp Leu Glu Gln Phe Ser Glu Lys Phe Tyr Cys Ser Cys 385 390 395 400 Tyr Ser Thr Leu Ser Cys Lys Glu Lys Ala Asp Val Lys Asp Thr Asp 405 410 415 Ala Val Asp Val Cys Ile Ala Asp Gly Val Cys Ile Asp Ala Phe Leu 420 425 430 Lys Pro Pro Met Glu Thr Glu Glu Pro Gln Ile Phe Tyr Asn Ala Ser 435 440 445 Pro Ser Thr 450 ＜210＞ 51 ＜211＞ 435 ＜212＞ PRT ＜213＞ Artificial sequence <220> ＜223＞ Hyal1 ＜400＞ 51 Met Ala Ala His Leu Leu Pro Ile Cys Ala Leu Phe Leu Thr Leu Leu 1 5 10 15 Asp Met Ala Gln Gly Phe Arg Gly Pro Leu Leu Pro Asn Arg Pro Phe 20 25 30 Thr Thr Val Trp Asn Ala Asn Thr Gln Trp Cys Leu Glu Arg His Gly 35 40 45 Val Asp Val Asp Val Ser Val Phe Asp Val Val Ala Asn Pro Gly Gln 50 55 60 Thr Phe Arg Gly Pro Asp Met Thr Ile Phe Tyr Ser Ser Gln Leu Gly 65 70 75 80 Thr Tyr Pro Tyr Tyr Thr Pro Thr Gly Glu Pro Val Phe Gly Gly Leu 85 90 95 Pro Gln Asn Ala Ser Leu Ile Ala His Leu Ala Arg Thr Phe Gln Asp 100 105 110 Ile Leu Ala Ala Ile Pro Ala Pro Asp Phe Ser Gly Leu Ala Val Ile 115 120 125 Asp Trp Glu Ala Trp Arg Pro Arg Trp Ala Phe Asn Trp Asp Thr Lys 130 135 140 Asp Ile Tyr Arg Gln Arg Ser Arg Ala Leu Val Gln Ala Gln His Pro 145 150 155 160 Asp Trp Pro Ala Pro Gln Val Glu Ala Val Ala Gln Asp Gln Phe Gln 165 170 175 Gly Ala Ala Arg Ala Trp Met Ala Gly Thr Leu Gln Leu Gly Arg Ala 180 185 190 Leu Arg Pro Arg Gly Leu Trp Gly Phe Tyr Gly Phe Pro Asp Cys Tyr 195 200 205 Asn Tyr Asp Phe Leu Ser Pro Asn Tyr Thr Gly Gln Cys Pro Ser Gly 210 215 220 Ile Arg Ala Gln Asn Asp Gln Leu Gly Trp Leu Trp Gly Gln Ser Arg 225 230 235 240 Ala Leu Tyr Pro Ser Ile Tyr Met Pro Ala Val Leu Glu Gly Thr Gly 245 250 255 Lys Ser Gln Met Tyr Val Gln His Arg Val Ala Glu Ala Phe Arg Val 260 265 270 Ala Val Ala Ala Gly Asp Pro Asn Leu Pro Val Leu Pro Tyr Val Gln 275 280 285 Ile Phe Tyr Asp Thr Thr Asn His Phe Leu Pro Leu Asp Glu Leu Glu 290 295 300 His Ser Leu Gly Glu Ser Ala Ala Gln Gly Ala Ala Gly Val Val Leu 305 310 315 320 Trp Val Ser Trp Glu Asn Thr Arg Thr Lys Glu Ser Cys Gln Ala Ile 325 330 335 Lys Glu Tyr Met Asp Thr Thr Leu Gly Pro Phe Ile Leu Asn Val Thr 340 345 350 Ser Gly Ala Leu Leu Cys Ser Gln Ala Leu Cys Ser Gly His Gly Arg 355 360 365 Cys Val Arg Arg Thr Ser His Pro Lys Ala Leu Leu Leu Leu Leu Asn Pro 370 375 380 Ala Ser Phe Ser Ile Gln Leu Thr Pro Gly Gly Gly Pro Leu Ser Leu 385 390 395 400 Arg Gly Ala Leu Ser Leu Glu Asp Gln Ala Gln Met Ala Val Glu Phe 405 410 415 Lys Cys Arg Cys Tyr Pro Gly Trp Gln Ala Pro Trp Cys Glu Arg Lys 420 425 430 Ser Met Trp 435

The above and other objects, features and other advantages of the present invention will be more clearly understood from the following detailed description with reference to the accompanying drawings. Figure 1A shows the particle size sizing chromatogram of trastuzumab in the stability test under the harsh conditions of 45°C. Figure 1B shows the variation in the purity of trastuzumab monomeric protein by formulation in a stability test under the harsh conditions of 45°C. Figures 2A and 2B show the results of measuring the protein aggregation temperature of formulations comprising trastuzumab and the novel PH20 variant HP46. Figure 3A is a weak cation exchange (WCX) chromatogram of trastuzumab in a stability test under severe conditions at 45°C. Figure 3B shows the change (%) in the relative amount of acidic variants in the formulations in the stability test under the harsh conditions of 45°C. Figure 3C shows the change (%) of the relative amount of the main peak in the formulation in the stability test under the harsh conditions of 45°C. Figure 3D shows the change (%) of the relative amount of basic variants in the formulations in the stability test under the harsh conditions of 45°C. Figure 4 shows the variation in the purity of trastuzumab monomeric protein in formulations 5-7 in a stability test under harsh conditions at 45°C. Figure 5A shows the change (%) in the relative amount of acidic variants in formulations 5-7 in the stability test under the harsh conditions of 45°C. Figure 5B shows the change (%) in the relative amount of the main peak in formulations 5-7 in the stability test under the harsh conditions of 45°C. Figure 5C shows the change (%) in the relative amount of basic variants in formulations 5-7 in the stability test under the harsh conditions of 45°C. Figure 6A shows Herceptin subcutaneous injection formulation (He Aiping SC), trastuzumab + wild type measured on Day 0 and Day 1 in a stability test under harsh conditions at 40°C Results of residual enzymatic activity of PH20 (HW2) and trastuzumab + PH20 variant HP46. Figure 6B shows Heaiping subcutaneous injection formulations, trastuzumab + wild-type PH20 (HW2) and trastuzumab + PH20 measured on days 0 and 1 in a stability test under harsh conditions at 45°C Results of residual enzyme activity of variant HP46. Figure 7 shows the particle size sieve chromatography analysis results of Formulations 8-10 in the 14-day stability test under severe conditions at 40°C. FIG. 8A shows the results of measuring the size change of protein particles using a dynamic light scattering (Dynamic light scattering, DLS) device. FIG. 8B shows the results of measuring protein aggregation temperature. Figure 9A shows a weak cation exchange (WCX) chromatogram of Formulation 8 in a stability test under harsh conditions at 40°C. Figure 9B shows the change (%) in the relative amount of acidic variants in formulations 8-10 in the stability test under the harsh conditions of 40°C. Figure 9C shows the change (%) in the relative amount of the main peak in formulations 8-10 in the stability test under the harsh conditions of 40°C. Figure 9D shows the change (%) in the relative amount of basic variants in formulations 8-10 in the stability test under the harsh conditions of 40°C. Figure 10 shows the change (%) of the relative enzyme activity of formulations 8-10 in the stability test under the harsh conditions of 40°C. Figure 11 shows the variation in the purity of trastuzumab monomers of Formulations 11-13 in a stability test under harsh conditions at 40°C. Figure 12A shows a weak cation exchange (WCX) chromatogram of formulation 11 in a stability test under harsh conditions at 40°C. Figure 12B shows the change (%) in the relative amount of acidic variants in formulations 11-13 in the stability test under the harsh conditions of 40°C. Figure 12C shows the change (%) in the relative amount of the main peak in formulations 11-13 in the stability test under the harsh conditions of 40°C. Figure 12D shows the change (%) of the relative amount of basic variants in formulations 11-13 in the stability test under the harsh conditions of 40°C. Figure 13 shows the change (%) of the relative enzyme activity of formulations 11-13 in the stability test under the harsh conditions of 40°C. Figure 14 shows the change in the purity of rituximab monomer in formulations 14-16 in a stability test under harsh conditions at 40°C. Figure 15 shows the change (%) of the relative enzyme activity of formulations 14-16 in the stability test under the harsh conditions of 40°C. Figure 16 shows the change in the relative enzyme activity of formulations 17 and 18 in the stability test under the harsh conditions of 40°C. Figure 17 shows the results of size sizing chromatography analysis of Formulations 19-22 at 40°C. Figure 18 shows the change in the relative enzyme activity of formulations 19-22 in the stability test under the harsh conditions of 40°C. Fig. 19 shows the change of the enzyme activity of recombinant human PH20 and HP46 according to the change of pH. Figure 20 shows a He Aiping subcutaneous injection product (He Aiping SC) and a biosimilar candidate for He Aiping subcutaneous injection (trastuzumab + HP46; Experimental results of pharmacokinetics of He Aiping SC BS), wherein He Aiping and He Aiping biosimilar candidates were injected at 18 mg/kg each, and the subcutaneous injection formulation contained 100 units of rHuPH20 and 100 units of HP46 (at pH 5.3).

Claims (27)

A pharmaceutical composition comprising: (a) a drug that is a protein drug, an antibody, a small molecule, an aptamer, an RNAi, an antisense RNA, or a cell therapy agent; and (b) a PH20 variant , comprising the substitution of amino acid residues of L354I and/or N356E of the wild-type PH20 of SEQ ID NO: 1, and further comprising an α-helical domain selected from the wild-type PH20 of SEQ ID NO: 1 and corresponding to the α One or more amino acid residue substitutions in one or more regions of the group consisting of the region connecting the domains of one of the helical domains, and wherein the α-helical domain of the wild-type PH20 of SEQ ID NO: 1 is α Helix 8 domain (S347 to C381), the linking domain of wild-type PH20 of SEQ ID NO: 1 is the linking domain (A333 to R346) between α-helix 7 and α-helix 8, and the PH20 variant has hyaluronic acid Enzyme activity. The pharmaceutical composition according to claim 1, wherein the α-helical domain and the region corresponding to the connecting domain of the α-helical domain are T341 to N363, T341 to I361, L342 to I361 of wild-type PH20 of SEQ ID NO: 1 , S343 to I361, I344 to I361, M345 to I361 or M345 to N363. The pharmaceutical composition as claimed in claim 1, wherein it is selected from the α-helix 8 domain (S347 to C381) of wild-type PH20 of SEQ ID NO: 1 and the connecting domain (A333) between α-helix 7 and α-helix 8 One or more regions of the group consisting of R346) are replaced by one or more amino acid residues of the amino acid sequence of the corresponding region of human hyaluronidase-1 (Hyal1). The pharmaceutical composition as claimed in item 1, wherein the PH20 variant is included in the group consisting of T341, L342, S343, I344, M345, S347, M348, K349, L352, L353, D355, E359, I361 and N363 Amino acid residue substitution at one or more positions of the group. The pharmaceutical composition according to claim 4, wherein the PH20 variant is selected from the group consisting of T341S, L342W, S343E, I344N, M345T, S347T, M348K, K349E, L352Q, L353A, D355K, E359D, I361T and N363G Group of one or more amino acid residue substitutions. The pharmaceutical composition according to claim 4, wherein the PH20 variant comprises amino acid residue substitutions of M345T, S347T, M348K, K349E, L352Q, L353A, L354I, D355K, N356E, E359D and I361T. The pharmaceutical composition according to claim 6, wherein the PH20 variant further comprises one or more substitutions of amino acid residues selected from the group consisting of T341S, L342W, S343E, I344N and N363G. The pharmaceutical composition according to claim 7, wherein the PH20 variant comprises any amino acid residue substitution selected from the following groups: (a) T341S, L342W, S343E, I344N, M345T, S347T, M348K, K349E (c3) , S347T, M348K, K349E, L352Q, L353A, L354I, D355K, N356E, E359D and I361T; (d) M345T, S347T, M348K, K349E, L352Q, L353A, L354I, D355K, N356E, E3619T and I361T; (d) ) I344N, M345T, S347T, M348K, K349E, L352Q, L353A, L354I, D355K, N356E, E359D and I361T; , E359D and I361T. The pharmaceutical composition according to claim 1, wherein the PH20 variant further comprises deletion of one or more amino acid residues at at least one of a carboxyl terminal and an amino terminal. The pharmaceutical composition according to claim 9, wherein in the PH20 variant, before the amino acid residues selected from the group consisting of M1 to P42, one or more missing amino acid residues. The pharmaceutical composition according to claim 10, wherein in the PH20 variant, before the amino acid residue L36, N37, F38, R39, A40, P41 or P42, a or multiple amino acid residues are missing. The pharmaceutical composition according to claim 9, wherein in the PH20 variant, after the amino acid residue selected from the group consisting of V455 to L509, one or more Amino acid residues are missing. The pharmaceutical composition according to claim 12, wherein in the PH20 variant, after the amino acid residue selected from the group consisting of V455 to S490, one or more Amino acid residues are missing. The pharmaceutical composition according to claim 13, wherein in the PH20 variant, at amino acid residues V455, C458, D461, C464, I465, D466, A467, F468, K470, P471, P472, M473, E474 , T475, E476, P478, I480, Y482, A484, P486, T488 or S490, one or more amino acid residues at the carboxy-terminus are deleted by excision. The pharmaceutical composition according to any one of claims 1 to 14, wherein the PH20 variant further comprises a message peptide at the amino terminal, the message peptide is derived from human hyaluronidase-1, human growth hormone or human serum albumin protein. The pharmaceutical composition according to claim 1, wherein the drug is the antibody, a soluble receptor or a fusion protein thereof. The pharmaceutical composition according to claim 1, wherein the antibody binds to one or more antigens selected from the group consisting of the following antigens: 4-1BB, integrin, amyloid beta, angiogenin, angiogenesis B cell activating factor (BAFF), B7-H3, complement 5, CCR4, CD3, CD4, CD6, CD11a, CD19, CD20, CD22, CD30, CD33, CD38, CD52, CD62, CD79b, CD80, CGRP, claudin-18, complement factor D, CTLA4, DLL3, EGF receptor, hemophilia factor, Fc receptor, FGF23, folate receptor, GD2, GM-CSF, HER2, HER3, interferon receptor, Interferon gamma, IgE, IGF-1 receptor, interleukin 1, interleukin 2 receptor, interleukin 4 receptor, interleukin 5, interleukin 5 receptor, interleukin 6, interleukin Interleukin 6 receptor, interleukin 7, interleukin 12/23, interleukin 13, interleukin 17A, interleukin 17 receptor A, interleukin 31 receptor, interleukin 36 receptor, LAG3 , LFA3, NGF, PVSK9, PD-1, PD-L1, RANK-L, SLAMF7, tissue factor, TNF, VEGF and one or more antigens of the group consisting of vWF. The pharmaceutical composition according to claim 1, wherein the antibody is one or more selected from the group consisting of the following antibodies: automilumab, natalizumab, etrolizumab, vedolizumab, bimagrumab, bapineuzumab, crenezumab, solanezumab, aducanumab, gantenerumab, AMG 780、MEDI 3617、nesvacumab、vanucizumab、evinacumab、tabalumab、lanalumab、belimumab、omburtamab、ravulizumab、eculizumab、mogamulizumab、otelixizumab、teplizumab、muromonab、tebentafusp、blinatumoma、REGN1979、ibalizumab、zanolimumab、itolizumab、efalizumab、inebilizumab、tafasitamab、 loncastuximab tesirine、ocrelizumab、ublituximab、obinutuzumab、ofatumumab、rituximab、tositumomab、ibritumomab tiuxetan、epratuzumab、inotuzumab ozogamicin、moxetumomab pasudotox、brentuximab vedotin、vadastuximab talirine、gemtuzumab ozogamicin、daratumumab、isatuximab、alemtuxumab、crizanlizumab、polatuzumab vedotin、galiximab、eptinezumab、 fremanezumab, galcanezumab, erenumab, zolbetuximab, lampalizumab, tremelimumab, zalifrelimab, ipilimumab, rovalpituzumab tesirine, cetuximab, depatuxizumab, zalutumumab, necitumumab, panitumumab, emicizumab, nipocalimab, rozanolixizumab, irburosumab, ximab soravtansine、dinutuximab、naxitamab、otilimab、margetuximab、pertuzumab、trastuzumab、trastuzumab deruxtecan、trastuzumab emtansine、trastuzumab duocarmazine、patritumab、anifrolumab、emapalumab、ligelizumab、omalizumab、dalotuzumab、figitumumab、teprotumumab、gebokizumab、canakinumab、daclizumab、basiliximab、dupilumab、 mepolizumab、reslizumab、benralizumab、clazakizumab、olokizumab、sirukumab、siltuximab、sarilumab、satralizumab、tocilizumab、REGN88、secukinumab、ustekinumab、briakinumab、lebrikizumab、tralokinumab、ixekizumab、bimekizumab、brodalumab、brazikumab、guselkumab、risankizumab、tildrakizumab、mirikizumab、nemolizumab、 spesolimab、relatlimab、narsoplimab、fasinumab、tanezumab、alirocumab、evolocumab、bococizumab、lambrolizumab、balstilimab、camrelizumab、cemiplimab、dostarlimab、prolgolimab、sintilimab、spartalizumab、tislelizumab、pembrolizumab、nivolumab、atezolizumab、avelumab、envafolimab、durvalumab、bintrafusp alpha、denosumab , elotuzumab, concizumab, marstacimab, infliximab, adalimumab, golimumab, certolizumab pegol, ozoralizumab, broluci zumab, ranibizumab, bevacizumab, faricimab, ramucirumab, and caplacizumab. The pharmaceutical composition according to claim 16, wherein the soluble receptor contained in the soluble receptor or an Fc fusion protein of the soluble receptor is selected from TNF-α soluble receptor, VEGF soluble receptor, CTLA-4 , a group consisting of soluble receptors for interleukin 1 and soluble receptors for LFA3. The pharmaceutical composition according to claim 19, wherein the Fc fusion protein of the soluble receptor is selected from the group consisting of etanercept, aflibercept, abatacept, belatacept, rilonacept and alefacept. The pharmaceutical composition as described in Claim 1 further comprises one or more selected from the group consisting of a buffer, a stabilizer and a surfactant. The pharmaceutical composition as described in claim 21, wherein the buffer solution comprises malate, formate, citrate, acetate, propionate, pyridine, piperidine, cacodylate, succinic acid Salt, 2-(N-𠰌linyl)ethanesulfonic acid (MES), histidine, tris(Tris), bis-(2-hydroxyethyl)amino tris Methane (bis-Tris), phosphate, ethanolamine, carbonate, piperazine-N,N'-bis(2-ethanesulfonic acid) (PIPES), imidazole, 1,3-bis(tri(hydroxymethyl)methane Amino)propane (BIS-TRIS propane), N,N-bis(2-hydroxyethyl)-2-aminoethanesulfonic acid (BES), 3-(N-𠰌linyl)propanesulfonic acid (MOPS ), hydroxyethylpiperone ethanesulfonic acid (HEPES), pyrophosphate and triethanolamine, one or more of the group consisting of; the stabilizer comprises a carbohydrate, a sugar or its hydrate, a sugar One or more of the group consisting of alcohol or its hydrate and an amino acid; and the surfactant comprises polyoxyethylene-sorbitol fatty acid ester, polyethylene-polypropylene glycol, polyoxyethylene-hard One or more nonionic surfactants in the group consisting of fatty acid ester, polyoxyethylene alkyl ether, polyoxyethylene-polyoxypropylene copolymer and sodium dodecylsulfonate (SDS). The pharmaceutical composition as described in claim 22, wherein the carbohydrate, the sugar or the sugar alcohol comprises trehalose or its hydrate, sucrose, saccharin, glycerin, butylene glycol, threitol, xylitol, Arabitol, ribitol, mannitol, sorbitol, galactitol, trehalitol, iditol, inositol, heptaheptitol, isomalt, maltitol, polysaccharide alcohol, cyclodextrin , one or more of the group consisting of hydroxypropyl cyclodextrin and glucose, and the amino acid comprises glutamic acid, glutamic acid, glycine, lysine, lysine and lysine Lysilysine, leucine, methionine, valine, serine, selenomethionine, citrulline, arginine, asparagine, aspartic acid, orimine , isoleucine, taurine, theanine, threonine, tryptophan, tyrosine, phenylalanine, proline, pyrrolysine, histidine and alanine one or more. The pharmaceutical composition according to claim 22, wherein the pharmaceutical composition comprises histidine buffer, trehalose and methionine, and the histidine buffer provides a pH of 5.5±2.0. The pharmaceutical composition according to claim 24, wherein the pharmaceutical composition comprises histidine buffer, trehalose, methionine and polysorbate, and the histidine buffer provides a pH of 5.5±2.0. The pharmaceutical composition as described in claim 25, wherein the pharmaceutical composition comprises the histidine buffer, 10-400 mM α, α-trehalose, 1-50 mM methionine and 0.0000001% (w/v ) to 0.5% (w/v) of polysorbate, the histidine buffer provides a pH of 5.5±2.0. An injection preparation for subcutaneous injection, comprising the pharmaceutical composition according to any one of Claims 1 to 26.

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