Pharmaceutical composition comprising human hyaluronidase PH20 variant and a drug for subcutaneous injection

Abstract

The present invention relates to a pharmaceutical composition comprising (a) a drug and (b) a human PH20 variant. The human PH20 variant included in the pharmaceutical composition according to the present invention comprises an alpha helix 8 sequence (S347-C381) selected from the wild-type human pH20 with the amino acid sequence SEQ ID NO:1 and an amino acid substitution at one or more positions of the connecting region (A333-R346) between alpha helix 7 and alpha helix 8, and is optionally cut at an amino acid located in the N-terminal or C-terminal region. In addition, the pharmaceutical composition according to the present invention may further include a pharmaceutically acceptable additive, particularly a stabilizer. Due to the advantages of human PH20 variants, the pharmaceutical composition according to the present invention can show the maximum therapeutic effect of the drug used therewith.

Description

Technical Field

[0001] This disclosure relates to a pharmaceutical composition comprising a human hyaluronidase PH20 variant having enhanced enzyme activity and thermal stability and one or more drugs; and a method of treating a disease using said pharmaceutical composition.

[0002] The pharmaceutical compositions according to this disclosure are preferably intended for subcutaneous injection. Background Technology

[0003] Medications requiring high or multiple doses (especially antibody drugs) are typically administered intravenously, a process that takes approximately 90 minutes or longer and involves additional preparation procedures, making it inconvenient and costly for patients, doctors, and healthcare professionals. In contrast, subcutaneous injection offers the advantage of immediate administration, but absorption is relatively lower compared to intravenous injection, and at doses of 3-5 mL or more, it can cause swelling and pain at the injection site due to slow absorption. For this reason, subcutaneous administration of protein therapies is usually limited to small solutions of 2 mL or less. However, when hyaluronidase is administered subcutaneously (or via subcutaneous injection) along with the therapeutic drug, hyaluronic acid distributed in the extracellular matrix is ​​hydrolyzed by hyaluronidase, reducing viscosity and increasing permeability in the subcutaneous region, thus allowing for the easy delivery of high or multiple doses of the drug into the body.

[0004] There are six types of hyaluronidase genes in humans: 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 membranes and acrosomal membranes. HyalPS1 is not expressed because it is a pseudogene. PH20 is an enzyme (EC 3.2.1.35) that cleaves the β-1,4 bond between N-acetylglucosamine and glucuronic acid (the sugar that makes up hyaluronic acid). The optimal pH for human hyaluronidase PH20 is 5.5, but it exhibits some activity even at pH 7-8, while other human hyaluronidases (including Hyal1) have an optimal pH of 3-4 and exhibit weak activity at pH 7-8. The pH of the human subcutaneous region is approximately 7.4, which is generally neutral; therefore, among the various types of hyaluronidase, PH20 is widely used in clinical applications. Examples of clinical applications of PH20 include subcutaneous injection of antibody therapeutic agents, used as an ocular relaxant and anesthetic additive in ophthalmic surgery; to increase the pathway for anticancer therapeutic agents to enter tumor cells by hydrolyzing hyaluronic acid in the extracellular matrix of tumor cells; and to promote the absorption of excess body fluids and blood in tissues.

[0005] Meanwhile, commercially available PH20 is currently in the form of extract from bovine or sheep testes. Examples include Amphadase® (bovine hyaluronidase) and Vitrase® (sheep hyaluronidase).

[0006] Bovine testicular hyaluronidase (BTH) is obtained by removing the signal peptide and 56 amino acids at the C-terminus from bovine wild-type PH20 during post-translational modification. BTH is also a glycoprotein and, based on its total amino acid composition, contains 5% mannose and 2.2% glucosamine (Borders and Raftery, 1968). When animal-derived hyaluronidase is repeatedly administered to humans in high doses, neutralizing antibodies can be produced, and other animal-derived biological materials, besides PH20, may cause allergic reactions as impurities. In particular, the use of bovine PH20 is restricted due to concerns about mad cow disease. To overcome these issues, research has been conducted on recombinant human PH20 protein.

[0007] Recombinant human PH20 protein has been reported to be expressed in yeast (Pichia pastoris), DS-2 insect cells, and animal cells (Chen et al., 2016; Hofinger et al., 2007). The recombinant PH20 protein produced in insect cells and yeast differs from human PH20 in its N-glycosylation pattern during post-translational modification.

[0008] 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 identified. Hyal1 consists of two domains: a catalytic domain and an EGF-like domain. The catalytic domain is in the form of (β / α)8, where the α-helix and β-chain (characteristic of secondary protein structures) are each repeated eight times (Chao et al., 2007). The EGF-like domain is completely conserved in variants of Hyal1 where the C-terminus is spliced ​​differently. Hyal1 shares 35.1% amino acid sequence identity with PH20, and the tertiary structure of PH20 has not yet been identified.

[0009] In structure / function studies of human PH20, the C-terminal region of PH20 has been found to be important for protein expression and enzyme activity. Specifically, it has been reported that terminating the C-terminus at amino acids 477-483 is important for enzyme expression and activity (Frost, 2007). The activity of full-length PH20 (amino acids 1-509) or PH20 variants with a C-terminus truncated at position 467 is only 10% or less of that of PH20 variants with a C-terminus truncated at one of positions 477 to 483 (Frost, 2007). Halozyme Therapeutics developed rHuPH20 (amino acids 36-482), a recombinant protein in which the C-terminus of mature PH20 is cleaved at Y482 (Bookbinder et al., 2006; Frost, 2007).

[0010] Meanwhile, although research is underway to develop various therapeutics using human PH20 via subcutaneous injection, the issue of low stability of human PH20 itself remains unresolved.

[0011] Against this technical background, the inventors of this disclosure have demonstrated that human PH20 variants (including substitutions of one or more amino acid residues in the α-helix 8 region (S347 to C381) and the linker region (A333 to R346) between α-helix 7 and α-helix 8 in the amino acid sequence of wild-type hyaluronidase PH20, wherein some amino acids located at the N-terminus and / or C-terminus of PH20 are cleaved) have high enzyme activity and thermostability, and have therefore filed a patent application (PCT / KR 2019 / 009215) for this purpose.

[0012] The inventors of this application have also confirmed that the PH20 variant according to this disclosure can be applied to pharmaceutical compositions or preparations containing drugs (e.g., antibody drugs, particularly high-dose anti-HER2 antibodies or immune checkpoint antibodies), and therefore the pharmaceutical compositions and preparations according to this disclosure (containing the PH20 variant and drugs, such as anti-HER2 antibodies or immune checkpoint antibodies) can be used for subcutaneous injection, and the activity of the drugs (such as antibody drugs) and the PH20 variant is very stable and can be maintained for a long time, thus completing this disclosure. Summary of the Invention

[0013] [Technical Issues]

[0014] Therefore, this disclosure is made in response to the above-mentioned problems, and one object of this disclosure is to provide a novel pharmaceutical composition comprising a PH20 variant having enhanced enzyme activity and thermal stability, and a drug, wherein the thermal stability and activity of the drug and the PH20 variant can be maintained for a long time; particularly a pharmaceutical composition suitable for subcutaneous injection.

[0015] Another object of this disclosure is to provide a method for treating a disease, the method comprising administering a pharmaceutical composition according to this disclosure to a subject in need of treatment.

[0016] [Technical Solution]

[0017] According to this disclosure, the above and other objectives can be achieved by providing a pharmaceutical composition comprising (a) a drug and (b) a PH20 variant.

[0018] The PH20 variant contained in the pharmaceutical composition according to this disclosure may comprise one or more amino acid residues selected from S343E, M345T, K349E, L353A, L354I, N356E and I361T of wild-type human PH20 having the amino acid sequence of SEQ ID NO: 1; and may further comprise one or more amino acid residues selected from one or more regions of the α-helix 8 region (S347 to C381) and / or the linker region (A333 to R346) between α-helix 7 and α-helix 8, wherein some amino acid residues located at the N-terminus and / or C-terminus are selectively cleaved.

[0019] The pharmaceutical compositions according to this disclosure may further comprise one or more of pharmaceutically acceptable additives, particularly buffers, stabilizers and surfactants.

[0020] The pharmaceutical compositions according to this disclosure can be used in the form of injectable preparations for subcutaneous injection. Attached Figure Description

[0021] The above and other objects, features and advantages of this disclosure will become clearer from the following detailed description taken in conjunction with the accompanying drawings, in which:

[0022] Figure 1 A shows a size exclusion chromatography chromatogram of trastuzumab in a stability test under harsh conditions of 45ºC, and Figure 1 B shows the changes in the purity of trastuzumab monomer protein according to the formulation during stability testing under harsh conditions of 45ºC.

[0023] Figure 2The results show the results of measuring the protein aggregation temperature of a formulation containing trastuzumab and the novel PH20 variant HP46;

[0024] Figure 3 A is the weak cation exchange (WCX) chromatogram of trastuzumab under harsh conditions of 45ºC. Figure 3 B shows the change (%) in the relative amount of acidic variants in the formulation during stability testing under harsh conditions at 45ºC. Figure 3 C shows the relative change (%) of the main peak of the formulation in the stability test under harsh conditions of 45ºC, and Figure 3 D shows the change (%) in the relative amount of basic variants in the formulation during stability testing under harsh conditions at 45ºC.

[0025] Figure 4 The changes in the purity of trastuzumab monomer protein in formulations 5-7 are shown in stability tests under harsh conditions of 45ºC.

[0026] Figure 5 A shows the change (%) in the relative amount of acidic variants in formulations 5-7 during stability testing under harsh conditions at 45ºC. Figure 5 B shows the change (%) in the relative amounts of the main peaks of formulations 5, 6, and 7 during stability testing under harsh conditions at 45ºC. Figure 5 C shows the change (%) in the relative amounts of basic variants of formulations 5, 6 and 7 during stability testing under harsh conditions at 45ºC.

[0027] Figure 6 A shows the results of residual enzyme activity measured on days 0 and 1 in a stability test under harsh conditions at 40ºC for Herceptin subcutaneous injection formulation (Herceptin SC), trastuzumab + wild-type PH20 (HW2), and trastuzumab + PH20 variant HP46. Figure 6 B shows the results of residual enzyme activity measured on days 0 and 1 in a stability test under harsh conditions of 45ºC for Herceptin subcutaneous injection formulation, trastuzumab + wild-type PH20 (HW2), and trastuzumab + PH20 variant HP46.

[0028] Figure 7 Size exclusion chromatography analysis results of formulations 8-10 were shown during a stability test conducted under harsh conditions of 40ºC for 14 days.

[0029] Figure 8 A shows the results of measuring protein particle size changes in formulations 8-10 using a DLS device, and Figure 8B shows the results of measuring the protein aggregation temperature;

[0030] Figure 9A The weak cation exchange (WCX) chromatogram of formulation 8 is shown in the stability test under harsh conditions of 40ºC. Figure 9B The changes (%) in the relative amounts of acidic variants in formulations 8-10 are shown in stability tests conducted under harsh conditions at 40ºC. Figure 9C The relative changes (%) of the main peak of formulations 8-10 in stability tests under harsh conditions of 40ºC are shown, and Figure 9D The relative changes (%) of basic variants in formulations 8-10 are shown in stability tests conducted under harsh conditions at 40ºC.

[0031] Figure 10 The changes (%) in relative enzyme activity of formulations 8-10 during stability testing under harsh conditions at 40ºC are shown.

[0032] Figure 11 The changes in the purity of trastuzumab monomers in formulations 11-13 are shown in stability tests under harsh conditions of 40ºC.

[0033] Figure 12A shows the weak cation exchange (WCX) chromatogram of formulation 11 in the stability test under harsh conditions of 40ºC; Figure 12B shows the change (%) of the relative amount of acidic variants in formulations 11-13 in the stability test under harsh conditions of 40ºC; Figure 12C shows the change (%) of the relative amount of the main peak in formulations 11-13 in the stability test under harsh conditions of 40ºC; and Figure 12D shows the change (%) of the relative amount of basic variants in formulations 11-13 in the stability test under harsh conditions of 40ºC.

[0034] Figure 13 The changes (%) in relative enzyme activity of formulations 11-13 during stability testing under harsh conditions at 40ºC are shown.

[0035] Figure 14 The changes in rituximab monomer purity in formulations 14-16 are shown in stability tests conducted under harsh conditions at 40ºC.

[0036] Figure 15 The changes in relative enzyme activity of formulations 14-16 under harsh conditions of 40ºC in stability tests are shown.

[0037] Figure 16 The changes in relative enzyme activity of formulations 17 and 18 are shown in stability tests under harsh conditions of 40ºC;

[0038] Figure 17 The results of size exclusion chromatography analysis of formulations 19-22 at 40ºC are shown.

[0039] Figure 18 The changes in relative enzyme activity of formulations 19-22 under harsh conditions of 40ºC in stability tests are shown.

[0040] Figure 19 The changes in enzyme activity caused by pH variations in recombinant human PH20 and HP46 are shown; and

[0041] Figure 20 The pharmacokinetic results of Herceptin subcutaneous product (Herceptin SC) and Herceptin subcutaneous biosimilar candidate (trastuzumab + HP46; Herceptin SC BS) in 9-week-old Sprague-Dawley rats are presented, wherein Herceptin and Herceptin biosimilar candidate were each injected at 18 mg / kg, and the subcutaneous formulation contained 100 units of rHuPH20 and 100 units of HP46 (at pH 5.3). Detailed Implementation

[0042] Unless otherwise defined, 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 pertains. Generally, the terms used herein are those well-known and commonly used in the art.

[0043] Embodiments of this disclosure relate to a pharmaceutical composition comprising (a) a drug and (b) a PH20 variant, and the pharmaceutical composition according to this disclosure can be used for the prevention or treatment of a disease, and is preferably used for subcutaneous injection.

[0044] The human PH20 variant contained in the pharmaceutical composition according to this disclosure has some amino acid residue substitutions in regions corresponding to the α-helix region and / or its linker region in the amino acid sequence of wild-type PH20 (having the amino acid sequence of SEQ ID NO: 1), preferably mature wild-type PH20 (having the sequence consisting of L36 to S490 in the amino acid sequence of SEQ ID NO: 1), preferably the α-helix 8 region (S347 to C381) and / or the linker region (A333 to R346) between α-helix 7 and α-helix 8, more preferably the amino acid region in T341 to N363, and most preferably T341 to I361, L342 to I361, S343 to I361, I344 to I361, M345 to I361 or M345 to N363.

[0045] In this disclosure, “mature wild-type PH20” means a protein containing the following amino acid residues: amino acid residues L36 to S490 of SEQ ID NO: 1, lacking M1 to T35 which form a signal peptide; and amino acid residues A491 to L509 of the amino acid sequence of wild-type PH20 having the sequence of SEQ ID NO: 1, which are not relevant to the substantive function of PH20.

[0046] Table 1. Amino acid sequence of wild-type PH20 (SEQ ID NO: 1)

[0047]

[0048] Specifically, the PH20 variant or fragment thereof contained in the pharmaceutical composition according to this disclosure comprises one or more mutations in wild-type PH20 having the sequence of SEQ ID NO: 1, amino acid residue substitutions preferably selected from S343E, M345T, K349E, L353A, L354I, N356E and I361T, and one or more amino acid residue substitutions most preferably selected from L354I and N356E.

[0049] In this disclosure, the term "PH20 variant" is intended to include mutations, preferred substitutions of amino acid residues in the sequence of wild-type human PH20; and deletions of some amino acid residues at the N-terminus and / or C-terminus along with such substitutions of amino acid residues, and is used in a substantially the same sense as the expression "PH20 variant or fragment thereof".

[0050] The inventors of this disclosure have demonstrated that novel PH20 variants or fragments thereof with enhanced enzyme activity and thermostability compared to wild-type PH20 can be provided based on previous research results in which the enzyme activity and protein aggregation temperature (Tagg) at neutral pH are increased when the amino acid sequences of the α-helix 8 region and the linker region between the α-helix 7 and α-helix 8 of human PH20 are partially replaced by the amino acid sequences of the α-helix 8 region and the linker region between the α-helix 7 and α-helix 8 of the highly hydrophilic Hyal1.

[0051] Therefore, the PH20 variant contained in the pharmaceutical composition according to this disclosure comprises one or more amino acid residues selected from S343E, M345T, K349E, L353A, L354I, N356E and I361T in the amino acid sequence of wild-type PH20 (having the amino acid sequence of SEQ ID NO: 1), preferably mature wild-type PH20 (having the sequence consisting of L36 to S490 in the amino acid sequence of SEQ ID NO: 1), preferably one or more amino acid residues selected from L354I and N356E.

[0052] In the region corresponding to the α-helix region and / or its linker region, preferably in the α-helix 8 region (S347 to C381) and / or in the linker region 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 are substituted.

[0053] In particular, in the PH20 variants included in the pharmaceutical compositions according to this disclosure, the α-helix 8 region (S347 to C381) of wild-type PH20, preferably mature wild-type PH20, and / or the linker region (A333 to R346) between α-helix 7 and α-helix 8 may be replaced by some amino acid residues in the amino acid sequence of the corresponding region of Hyal1 having the sequence SEQ ID NO: 51 (see Tables 2 and 3), but this disclosure is not limited thereto.

[0054] Table 2. Amino acid sequence of wild-type Hyal1 (SEQ ID NO: 51)

[0055]

[0056] Table 3. Comparison of α-helix and amino acid sequences between PH20 and Hyal1

[0057]

[0058] More specifically, the PH20 variant or fragment thereof contained in the pharmaceutical composition according to this disclosure preferably includes amino acid residues of L354I and / or N356E in the amino acid sequence of wild-type PH20, preferably mature wild-type PH20.

[0059] Furthermore, it is preferred that amino acid residue substitutions be further included at one or more positions selected from T341 to N363, particularly at one or more positions selected from T341, L342, S343, I344, M345, S347, M348, K349, L352, L353, D355, E359, I361, and N363, but this disclosure is not limited thereto, and

[0060] More preferably, it further comprises one or more amino acid residues selected from T341S, L342W, S343E, I344N, M345T, S347T, M348K, K349E, L352Q, L353A, D355K, E359D, I361T and N363G, but this disclosure is not limited thereto.

[0061] Preferably, the PH20 variant or fragment thereof included in the pharmaceutical composition according to this disclosure may contain amino acid residue substitutions selected from M345T, S347T, M348K, K349E, L352Q, L353A, L354I, D355K, N356E, E359D, and I361T.

[0062] It may further include one or more amino acid residues selected from T341S, L342W, S343E, I344N and N363G as substitutions, but this disclosure is not limited thereto.

[0063] More preferably, the PH20 variant or fragment thereof contained in the pharmaceutical composition according to this disclosure may contain, but is not limited to, any substitution selected from the group consisting of:

[0064] (a) T341S, L342W, S343E, I344N, M345T, S347T, M348K, K349E, L352Q, L353A, L354I, D355K, N356E, E359D and I361T;

[0065] (b) L342W, S343E, I344N, M345T, S347T, M348K, K349E, L352Q, L353A, L354I, D355K, N356E, E359D and I361T;

[0066] (c) M345T, S347T, M348K, K349E, L352Q, L353A, L354I, D355K, N356E, E359D and I361T;

[0067] (d) M345T, S347T, M348K, K349E, L352Q, L353A, L354I, D355K, N356E, E359D, I361T and N363G;

[0068] (e) I344N, M345T, S347T, M348K, K349E, L352Q, L353A, L354I, D355K, N356E, E359D and I361T; and

[0069] (f) S343E, I344N, M345T, S347T, M348K, K349E, L352Q, L353A, L354I, D355K, N356E, E359D and I361T.

[0070] In this disclosure, expressions such as “S347”, which are described by a one-letter amino acid residue code along with a number, refer to the amino acid residue at the corresponding position in the amino acid sequence of SEQ ID NO: 1.

[0071] For example, "S347" means that the amino acid residue at position 347 in the amino acid sequence of SEQ ID NO: 1 is serine. Additionally, "S347T" means that the serine residue at position 347 in SEQ ID NO: 1 is replaced by threonine.

[0072] According to this disclosure, the PH20 variant contained in the pharmaceutical composition is interpreted as including a variant in which an amino acid residue at a specific amino acid residue position is conservatively substituted.

[0073] As used herein, the term “conservative substitution” refers to a modification of the PH20 variant in which one or more amino acids are replaced by amino acids with similar biochemical properties, the latter of which do not cause a loss of the biological or biochemical function of the corresponding PH20 variant.

[0074] "Conservative amino acid substitution" is a substitution in which an amino acid residue is replaced by an amino acid residue having a similar side chain. Families of amino acid residues with similar side chains have been defined and are well known in the art. These families include amino acids with basic side chains (e.g., lysine, arginine, and histidine), amino acids with acidic side chains (e.g., aspartic acid and glutamic acid), amino acids with uncharged polar side chains (e.g., glycine, asparagine, glutamine, serine, threonine, tyrosine, and cysteine), amino acids with nonpolar side chains (e.g., alanine, valine, leucine, isoleucine, proline, phenylalanine, methionine, and tryptophan), amino acids with β-branched side chains (e.g., threonine, valine, and isoleucine), and amino acids with aromatic side chains (e.g., tyrosine, phenylalanine, tryptophan, and histidine).

[0075] It is expected that the PH20 variant contained in the pharmaceutical composition according to this disclosure will retain its activity despite having conserved amino acid substitutions.

[0076] Furthermore, the PH20 variant or fragment thereof contained in the pharmaceutical composition according to this disclosure is interpreted as including the following PH20 variant or fragment thereof having substantially the same function and / or effect as the PH20 variant or fragment thereof according to this disclosure, and having at least 80% or 85%, preferably at least 90%, more preferably at least 95%, and most preferably at least 99% amino acid sequence homology with the PH20 variant or fragment thereof according to this disclosure.

[0077] Compared to mature wild-type PH20, the PH20 variant according to this disclosure exhibits increased expression levels and increased protein refolding rate in animal cells, resulting in improved thermal stability. Furthermore, despite the improved thermal stability, the PH20 variant exhibits enzymatic activity exceeding or similar to that of mature wild-type PH20.

[0078] Furthermore, it is known that enzyme activity decreases when certain amino acids (such as S490) at the C-terminus of mature wild-type PH20 are additionally cleaved. However, the PH20 variant according to this disclosure exhibits improved thermostability and increased or similar enzyme activity compared to mature wild-type PH20, even with additional cleavage sequences at the C-terminus of mature wild-type PH20. Additionally, the PH20 variant maintains its enzyme activity when cleaved from up to five amino acid residues at the N-terminus, suggesting that residues starting at P41 at the N-terminus play a crucial role in protein expression and enzyme activity.

[0079] Therefore, the PH20 variant contained in the pharmaceutical composition according to this disclosure includes substitutions of some amino acid residues in the α-helix 8 region (S347 to C381) of wild-type PH20 and / or in the linker region (A333 to R346) between α-helix 7 and α-helix 8, and further includes deletions of some amino acid residues at the C-terminus and / or N-terminus, but this disclosure is not limited thereto.

[0080] In one embodiment, the PH20 variant included in the pharmaceutical composition according to this disclosure may comprise a deletion of some amino acid residues at the N-terminus resulting from a cleavage selected from SEQ ID NO: The amino acid sequence of 1 is preceded by amino acid residues M1 to P42 at the N-terminus, preferably before amino acid residues L36, N37, F38, R39, A40, P41, or P42; and / or by the deletion of some amino acid residues at the C-terminus caused by the following cleavage, said cleavage being after amino acid residues selected from V455 to W509 at the C-terminus, preferably after amino acid residues selected from V455 to S490, and most preferably after 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.

[0081] The statement "cleavage before L36, N37, F38, R39, A40, P41, or P42 at the N-terminus" means cleaving and removing all amino acid residues immediately preceding L36 (M1 to T35), immediately preceding N37 (M1 to L36), immediately preceding F38 (M1 to N37), immediately preceding R39 (M1 to F38), immediately preceding A40 (M1 to R39), immediately preceding P41 (M1 to A40), and immediately preceding P42 (M1 to P41). The statement "cleavage before M1 at the N-terminus of SEQ ID NO: 1" means no cleavage occurs at the N-terminus.

[0082] Additionally, the statement "cleavage following 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 C-terminus" means cleaving and removing the amino acid residues following V455, C458, D461, C464, I465, D466, A467, F468, K470, P472, M473, E474, T475, E476, P478, I480, Y482, A484, P486, T488, or S490 in the sequence of SEQ ID NO: 1. For example, cutting after S490 means cutting between S490 and A491.

[0083] Preferably, the human PH20 variant contained in the pharmaceutical composition according to this disclosure may have an amino acid sequence selected from the amino acid sequences of SEQ ID NO: 5 to 50, more preferably having the amino acid sequence of SEQ ID NO: 44, but this disclosure is not limited thereto. In the PH20 variant constructed according to a specific embodiment of this disclosure, the substituted or cleaved amino acid sequences are shown in Table 4 below.

[0084] Table 4. Amino acid sequences and substitution / cleavage characteristics of the PH20 variants according to this disclosure.

[0085]

[0086]

[0087]

[0088]

[0089]

[0090]

[0091]

[0092]

[0093]

[0094]

[0095]

[0096]

[0097]

[0098]

[0099]

[0100]

[0101]

[0102]

[0103]

[0104]

[0105]

[0106]

[0107]

[0108]

[0109] Meanwhile, previous studies have reported that the enzyme activity of wild-type PH20 varies depending on the cleavage position of the amino acid residues located at the C-terminus. However, in this disclosure, the specific α-helix forming the secondary structure of PH20 is replaced by the α-helix of other human hyaluronidases, thereby constructing PH20 variants with higher stability than wild-type PH20. In these variants, the interaction between the substituted α-helix domain of PH20 and other secondary structures exhibits a different pattern than that of wild-type PH20, thus the variants possess certain or higher levels of enzyme activity regardless of the cleavage position at the C-terminus.

[0110] In addition, this disclosure attempts to increase the expression of recombinant PH20 protein by replacing the original signal peptide of human PH20 with the signal peptide of other proteins that exhibit high protein expression levels in animal cells.

[0111] Therefore, in another embodiment, the PH20 variant included in the pharmaceutical composition according to this disclosure may contain a signal peptide derived from human hyaluronidase-1 (Hyal1), human growth hormone, or human serum albumin at its N-terminus instead of the wild-type PH20 signal peptide composed of M1 to T35, and preferably may contain a human growth hormone-derived signal peptide having the amino acid sequence MATGSRTSLLLAFGLLCLPWLQEGSA according to SEQ ID NO: 2, a human serum albumin-derived signal peptide having the amino acid sequence MKWVTFISLLFLFSSAYS according to SEQ ID NO: 3, or a human Hyal1-derived signal peptide having the amino acid sequence MAAHLLPICALFLTLLDMAQG according to SEQ ID NO: 4, but this disclosure is not limited thereto.

[0112] Table 5. Amino acid sequences of signal peptides from human growth hormone, human serum albumin, or human Hyal1.

[0113]

[0114] Among the PH20 variants included in the pharmaceutical compositions according to this disclosure, the variant having a 6xHis tag attached to its C-terminus is referred to as HM, while the variant without a 6xHis tag is referred to as HP. Additionally, mature wild-type PH20 (L36-S490) having a 6xHis tag attached to its C-terminus is referred to as WT, while mature wild-type PH20 (L36 to Y482) without a 6xHis tag, wherein the C-terminus is cleaved after Y482, is referred to as HW2.

[0115] HP46 (SEQ ID NO: 44) is a human PH20 variant obtained by modeling the protein structure using Hyal1 (PDB ID: 2PE4) (Chao et al., 2007), which has a known protein tertiary structure and is human hyaluronidase. The amino acid sequence of α-helix 8 and the linker region between α-helix 7 and α-helix 8 is then substituted with the amino acid sequence of Hyal1, and the N-terminus is cleaved at F38 and the C-terminus is cleaved after F468. Specifically, α-helix 8 is located outside the PH20 protein tertiary structure and interacts less with adjacent α-helices or β chains compared to other α-helices. Generally, there is a trade-off between enzyme activity and thermostability; therefore, higher protein thermostability corresponds to lower enzyme activity, and thermostability tends to decrease as enzyme activity increases due to increased protein structural flexibility. However, the specific activity of HP46, measured by turbidity determination at pH 7.0, was approximately 46 units / µg, which was evaluated as approximately twice the specific activity of wild-type PH20, which had a specific activity of approximately 23 units / µg.

[0116] The thermal stability of proteins can be evaluated based on the melting temperature Tm (at which 50% of the protein's tertiary structure denatures) and the aggregation temperature Tagg (at which proteins aggregate). Generally, the aggregation temperature of proteins tends to be lower than their melting temperature. The α-helix 8 of Hyal1 is more hydrophilic than that of PH20. The substituted α-helix 8 of Hyal1 increases the surface hydrophilicity of HP46, resulting in a delayed aggregation effect due to hydrophobic interactions, thus achieving an aggregation temperature of 51ºC, which is observed to be 4.5ºC higher than that of wild-type PH20 (46.5ºC).

[0117] HP46 is a variant in which amino acid residues in α-helix 8 and the linker region between α-helix 7 and α-helix 8 are substituted, with T341 being replaced by serine. When amino acid residue 341 is threonine, the enzyme activity is similar to that of wild-type PH20, but after substitution with serine, the enzyme activity is increased by approximately 2-fold. Furthermore, it can be confirmed that even in substrate gel assays, the resulting variant hydrolyzes 5 to 6 times more hyaluronic acid than wild-type PH20. Substrate gel assays involve protein denaturation and refolding processes, meaning that HP46 exhibits enhanced tertiary structure refolding and recovery compared to wild-type PH20.

[0118] The amount of the PH20 variant in the pharmaceutical composition according to this disclosure is at least 50 units / mL, preferably in the range of 100 units / mL to 20,000 units / mL, more preferably in the range of about 150 units / mL to about 18,000 units / mL, still more preferably in the range of 1,000 units / mL to 16,000 units / mL, and most preferably in the range of 1,500 units / mL to 12,000 units / mL.

[0119] Examples of pharmaceuticals included in the pharmaceutical compositions according to this disclosure include, but are not limited to, protein drugs, antibody drugs, small molecules, aptamers, RNAi, antisenses, and cell therapy agents (such as chimeric antigen receptor (CAR)-T or CAR-natural killer (NK) cells), and may include not only currently commercially available drugs, but also drugs in clinical trials or under development.

[0120] As a drug, protein drugs or antibody drugs are preferred.

[0121] The “protein drug” included in the pharmaceutical composition according to this disclosure is a drug composed of amino acids and thus exhibiting a therapeutic or preventive effect on disease through the activity of proteins; a drug composed of proteins other than antibody drugs; and may be selected from cytokines, therapeutic enzymes, hormones, soluble receptors and their fusion proteins, insulin or analogues thereof, bone morphogenetic protein (BMP), erythropoietin and serum-derived proteins, but this disclosure is not limited thereto.

[0122] The cytokines contained in the pharmaceutical compositions according to this disclosure may be selected from interferon, interleukin, colony-stimulating factor (CSF), tumor necrosis factor (TNF) and tissue growth factor (TGF), but this disclosure is not limited thereto.

[0123] Therapeutic enzymes may include, but are not limited to, β-glucocerebrosidase and agalsidase β.

[0124] The soluble receptor included in the pharmaceutical composition according to this disclosure is the extracellular domain of a receptor, and its fusion protein is a protein in which the Fc region of an antibody is fused with the soluble receptor. A soluble receptor is a soluble form of a disease-associated ligand-binding receptor, and examples include forms in which the Fc region is fused with a TNF-α soluble receptor (e.g., products containing the ingredient etanercept and similar forms), forms in which the Fc region is fused with a VEGF soluble receptor (e.g., products containing the ingredient alefacept and similar forms), forms in which the Fc region is fused with a CTLA-4 receptor (e.g., products containing the ingredients abatacept or beraccept and similar forms), forms in which the Fc region is fused with an interleukin-1 soluble receptor (e.g., products containing the ingredient linascept and similar forms), and forms in which the Fc region is fused with an LFA3 soluble receptor (e.g., products containing the ingredient alefacept and similar forms), but this disclosure is not limited thereto.

[0125] The hormones contained in the pharmaceutical compositions according to this disclosure refer to hormones or analogues thereof injected into the body for the treatment or prevention of diseases caused by hormone deficiency, and examples of said hormones or analogues include, but are not limited to, human growth hormone, estrogen, and progesterone.

[0126] The serum-derived proteins contained in the pharmaceutical compositions according to this disclosure are proteins present in plasma, and include both proteins extracted from plasma and recombinant proteins produced therefrom, and examples may include, but are not limited to, fibrinogen, von Willebrand factor, albumin, thrombin, factor II (FII), factor V (FV), factor VII (FVII), factor IX (FIX), factor X (FX), and factor XI (FXI).

[0127] The antibody drug contained in the pharmaceutical composition according to this disclosure may be a monoclonal antibody drug or a polyclonal antibody drug.

[0128] According to this disclosure, a monoclonal antibody drug comprises a monoclonal antibody and a protein containing a monoclonal antibody fragment capable of specifically binding to an antigen associated with a specific disease. Monoclonal antibodies also include bispecific antibodies, and proteins containing monoclonal antibodies or fragments thereof conceptually include antibody-drug conjugates (ADCs).

[0129] Examples of antigens associated with specific diseases include 4-1BB, integrin, β-amyloid, angiopoietin (angiopoietin 1 or 2), angiopoietin analogue 3, 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, tight junction protein-18, complement factor D, CTLA4, DLL3, EGF receptor, hemophilia factor, Fc receptor, FGF23, folate receptor, GD2, and GM. -CSF, HER2, HER3, interferon receptor, interferon-γ, IgE, IGF-1 receptor, interleukin-1, interleukin-2 receptor, interleukin-4 receptor, interleukin-5, interleukin-5 receptor, interleukin-6, 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, VEGF receptor and von Willebrand factor (vWF), but this disclosure is not limited thereto.

[0130] The following are, but are not limited to, proteins, including monoclonal antibodies or monoclonal antibody fragments targeting antigens associated with specific diseases:

[0131] Utomilumab is an anti-4-1BB antibody.

[0132] Natalizumab, etorizumab, vedolizumab, and bigromab are antibodies targeting integrin.

[0133] Bapineuzumab, crenezumab, solanezumab, aducanumab, and ganteneerumab are antibodies targeting β-amyloid.

[0134] Antibodies against angiopoietin, such as AMG780 against angiopoietin 1 and 2, MEDI 3617 and Nescalumab against angiopoietin 2, and vancomycin as a bispecific antibody against angiopoietin 2 and VEGF.

[0135] Eviacumab is an antibody targeting angiopoietin analogue 3.

[0136] Tabalumab, lanalumab, and belimumab are antibodies targeting B cell activating factor (BAFF).

[0137] omburtamab is an antibody targeting B7-H3.

[0138] ravulizumab and eculizumab are antibodies targeting complement 5;

[0139] Mogamulizumab is an antibody targeting CCR4.

[0140] Osteizumab, 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.

[0141] Ibacillinumab and zanolimumab are antibodies targeting CD4.

[0142] Itrazumab is an antibody targeting CD6.

[0143] efalizumab is an antibody targeting CD11a.

[0144] Inbilizumab, tafasitamab, and loncastuximab tesirine are antibodies against CD19 (the loncastuximab is an ADC).

[0145] Oribitusumab, ublituximab, obbituzumab, oflamumab, rituximab, tosimomab, and ibritumomab tiuxetan are antibodies against CD20 (the latter being an ADC).

[0146] Epratuzumab, inotuzumabozogamicin (the inotuzumab being an ADC), and moxetumomab pasudotox are antibodies against CD22.

[0147] Brentuximab vedotin is an ADC targeting CD30.

[0148] Tadalirine-vadastuximab talirine and gemtuzumab ozogamicin are ADCs targeting CD33.

[0149] Daratumumab and isatuximab are antibodies targeting CD38.

[0150] alemtuzumab is an antibody targeting CD52;

[0151] Crizanlizumab is an antibody targeting CD62.

[0152] Porcuzumab vedotin is an ADC targeting CD79b.

[0153] Galiximab is an antibody targeting CD80;

[0154] Eptinezumab, fremanezumab, galcanezumab, and erenumab are antibodies against CGRP.

[0155] Zoloximab is an antibody targeting tight junction protein-18;

[0156] Lanpamumab is an antibody targeting complement factor D.

[0157] Trimemomab, zalifrelimab, and ipilimumab are antibodies targeting CTLA4.

[0158] Rovapituzumab tesirine is an ADC targeting DLL3.

[0159] Cetuximab, depatuxizumab, zalumumab, necitumumab, and panitumumab are antibodies targeting the EGF receptor.

[0160] emicizumab is a bispecific antibody targeting coagulation factor IX and factor X (which are hemophilia factors).

[0161] Nipocalimab and rozanolixizumab are antibodies targeting the Fc receptor.

[0162] Burosumab is an antibody targeting FGF23.

[0163] Farletuzumab, an antibody targeting the folate receptor, and mirvetuximab soravtansine, an ADC targeting the folate receptor.

[0164] Dinutuximab and naxitamab are antibodies targeting GD2.

[0165] Otelimab is an antibody targeting GM-CSF;

[0166] Margetuximab, pertuzumab, and trastuzumab are antibodies against HER2, while trastuzumab deruxtecan, trastuzumab emtansine, and trastuzumab duocarmazine are ADCs against HER2.

[0167] Patritumab is an antibody targeting HER3.

[0168] Anifrolumab is an antibody that targets the interferon receptor.

[0169] Emapalumab is an antibody targeting interferon-gamma.

[0170] Ligozumab and omalizumab are antibodies targeting IgE.

[0171] Dalotuzumab, figitumumab, and teprotumumab are antibodies targeting the IGF-1 receptor.

[0172] Gebokizumab and canakinumab are antibodies against interleukin-1.

[0173] Dacizumab and baliximab are antibodies targeting the interleukin-2 receptor;

[0174] Dupilumab is an antibody targeting the interleukin-4 receptor.

[0175] Mepozumab and Rosumab are antibodies targeting interleukin-5;

[0176] Benalizumab is an antibody targeting the interleukin-5 receptor.

[0177] Clagizumab, ologiizumab, slucurumab, and stoximab are antibodies targeting interleukin-6.

[0178] Sarralizumab, satralizumab, tocilizumab, and REGN88 are antibodies targeting the interleukin-6 receptor.

[0179] Secukinumab is an antibody targeting interleukin-7;

[0180] Utecumab and briakinumab are antibodies against interleukin-12 / 23.

[0181] Lebrikizumab and tralokinumab are antibodies against interleukin-13.

[0182] Ixekizumab and bimekizumab are antibodies against interleukin-17A.

[0183] Brodalumab is an antibody targeting interleukin-17 receptor A.

[0184] Antibodies against interleukin-23 include brazikumab, guselkumab, risankizumab, tildrakizumab, and mirikizumab.

[0185] Nemozumab is an antibody targeting the interleukin 31 receptor;

[0186] Pesolimab is an antibody targeting the interleukin-36 receptor.

[0187] Relatlimab is an antibody targeting LAG3.

[0188] Narsoplimab is an antibody targeting NSP2.

[0189] Faximab and tanezumab are antibodies against NGF.

[0190] Alikumab, evokumab, and bococizumab are antibodies targeting PVSK9.

[0191] Antibodies targeting PD-1 include lambrolizumab, balstilimab, camrelizumab, cemiplimab, dostarlimab, prolgolimab, shintilimab, spartalizumab, tislelizumab, pembrolizumab, and nivolumab.

[0192] Atezolizumab, avelumumab, envafolimab, and durvalumab are antibodies against PD-L1, and bintrafusp α is a bispecific antibody against TGFβ and PD-L1.

[0193] Denosumab, an antibody targeting RANK-L;

[0194] Elozizumab, an antibody targeting SLAMF7;

[0195] Concizumab and marstacimab are antibodies against tissue factor.

[0196] Antibodies against TNF, particularly TNFα, include infliximab, adalimumab, golimumab, the antibody fragment certolizumab pegol, and ozoralizumab (ozoralizumab is a bispecific antibody against TNF and albumin).

[0197] Antibodies against VEGF include brolucizumab, ranibizumab, bevacizumab, and faricimab (faricimab is a bispecific antibody against VEGF and Ang2).

[0198] Ramucirumab, an antibody targeting the VEGF receptor; and

[0199] Caplacizumab is an antibody targeting vWF.

[0200] Meanwhile, overexpression of human epidermal growth factor receptor 2 (HER2), which promotes cell division, has been observed in approximately 20%-25% of breast cancer patients. HER2-overexpressing breast cancer progresses rapidly, is aggressive, and responds less to chemotherapy compared to HER2-low-expressing breast cancer, thus having a poor prognosis. Trastuzumab is a monoclonal antibody drug targeting HER2. It specifically binds to HER2 on the surface of HER2-overexpressing cancer cells to inhibit signal transduction of cell replication and proliferation, thereby slowing tumor progression. Trastuzumab was approved by the U.S. Food and Drug Administration (FDA) for the treatment of breast cancer in 1998 and by the Korean Food and Drug Administration (KFDA) in 2003. Subsequently, the efficacy of trastuzumab has also been recognized in gastric cancer overexpressing HER2, and it has been used as a therapeutic agent for gastric cancer.

[0201] Roche's intravenous formulation of Herceptin (trade name: Herceptin) consists of 440 mg of trastuzumab as the main ingredient, which is lyophilized and mixed with saline before being injected intravenously. On the other hand, the subcutaneous formulation of trastuzumab (trade name: Herceptin SC) is a 5 mL liquid formulation containing 600 mg (120 mg / mL) of trastuzumab as the main ingredient, and includes 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.

[0202] Herceptin subcutaneous injection formulation has a shelf life of 21 months. Trastuzumab intravenous injection formulation is lyophilized and has a shelf life of 30 months, but trastuzumab subcutaneous injection formulation is liquid and has a short shelf life of 21 months. Therefore, it can be estimated that the stability of one or more of trastuzumab and recombinant human hyaluronidase PH20 in liquid formulations is limited.

[0203] In this context, given the characteristics of the PH20 variant according to this disclosure, wherein the PH20 variant not only has increased enzyme activity but also a measured high protein aggregation temperature compared to wild-type human hyaluronidase PH20 and recombinant human PH20 obtained from Halozyme, thus exhibiting improved thermal stability, the shelf life of the subcutaneous injection preparation is set to be long-term, preferably 21 months or longer.

[0204] The content of the antibody drug in the pharmaceutical composition according to this disclosure may be in the following ranges: 5 mg / mL to 500 mg / mL, preferably 20 mg / mL to 200 mg / mL, more preferably 100 mg / mL to 150 mg / mL, and most preferably 120 ± 18 mg / mL, for example, about 110 mg / mL, about 120 mg / mL or about 130 mg / mL.

[0205] The polyclonal antibodies contained in the pharmaceutical compositions according to this disclosure are preferably serum antibodies extracted from serum, such as immunoglobulins, but not limited thereto.

[0206] In the case of small molecule compounds, any drug requiring rapid preventative or therapeutic effects can be used without restriction. For example, morphine-based analgesics can be used (Thomas et al., 2009). Additionally, when used as a therapeutic agent for tissue necrosis induced by anticancer drugs, small molecule compounds can be used alone or in combination with antidotes such as vinca alkaloids and taxanes (Kreidieh et al., 2016).

[0207] The pharmaceutical compositions according to this disclosure may further comprise one or more selected from buffers, stabilizers, and surfactants.

[0208] The buffer solutions contained in the compositions according to this disclosure may be used without limitation, provided that they achieve a pH of 4 to 8, preferably 5 to 7, and the buffer solution is preferably selected from one or more of the following: malate, formate, citrate, acetate, propionate, pyridine, piperazine, carcoate, succinate, 2-(N-morpholino)ethanesulfonic acid (MES), histidine, Tris, bis-Tris, phosphate, ethanolamine, carbonate, piperazine-N,N'-bis(2-ethanesulfonic acid) (PIPES), imidazole, BIS-TRIS propane, N,N-bis(2-hydroxyethyl)-2-aminoethanesulfonic acid (BES), 3-(N-morpholino)propanesulfonic acid (MOPS), hydroxyethylpiperazine ethanesulfonic acid (HEPES), pyrophosphate, and triethanolamine, more preferably histidine buffers, such as L-histidine / HCl, but not limited thereto.

[0209] The concentration of the buffer solution can be in the following ranges: 0.001 mM to 200 mM, preferably 1 mM to 50 mM, more preferably 5 mM to 40 mM, and most preferably 10 mM to 30 mM.

[0210] Stabilizers in the compositions according to this disclosure may be used without limitation, provided that they are generally used in the art for the purpose of stabilizing proteins, and preferably, the stabilizers may be, for example, one or more selected from: carbohydrates, sugars or their hydrates, sugar alcohols or their hydrates, and amino acids.

[0211] The carbohydrate, sugar, or sugar alcohol used as a stabilizer may be selected from one or more of the following: trehalose or its hydrate, sucrose, saccharin, glycerol, erythritol, threitol, xylitol, arabinitol, ribitol, mannitol, sorbitol, galactitol, fucitol, idutol, inositol, heptaheptanol, isomaltitol, maltitol, polyglucitol, cyclodextrin, hydroxypropyl cyclodextrin, and glucose, but is not limited thereto.

[0212] The amino acid may be selected from one or more of the following: glutamine, glutamic acid, glycine, lysine, lysine dimer, leucine, methionine, valine, serine, selenomethionine, citrulline, arginine, asparagine, aspartic acid, ornithine, isoleucine, taurine, theanine, threonine, tryptophan, tyrosine, phenylalanine, proline, pyrrolidone, histidine, and alanine, but is not limited thereto.

[0213] The concentration of sugar or sugar alcohol used as a stabilizer in the pharmaceutical composition according to this disclosure can be in the following ranges: 0.001 mM to 500 mM, preferably 100 mM to 300 mM, more preferably 150 mM to 250 mM, and most preferably 180 mM to 230 mM, and particularly can be about 210 mM.

[0214] In addition, the concentration of the amino acid used as a stabilizer in the pharmaceutical composition according to this disclosure can be in the following range: 1 mM to 100 mM, preferably 3 mM to 30 mM, more preferably 5 mM to 25 mM, and most preferably 7 mM to 20 mM, and particularly in the range of 8 mM to 15 mM.

[0215] The compositions according to the invention may further contain surfactants.

[0216] Preferably, the surfactant can be a nonionic surfactant, such as polyoxyethylene-sorbitan fatty acid ester (polysorbate or Tween), polyethylene-polypropylene glycol, polyoxyethylene-stearate, polyoxyethylene alkyl ether, such as polyoxyethylene monolauryl ether, alkylphenyl polyoxyethylene ether [Triton-X], and polyoxyethylene-polyoxypropylene copolymer [poloxam and Plonic] and sodium dodecyl sulfate (SDS), but is not limited thereto.

[0217] More preferably, polysorbate can be used. The polysorbate can be polysorbate 20 or polysorbate 80, but is not limited thereto.

[0218] The concentration of the nonionic surfactant in the pharmaceutical composition according to this disclosure may be in the following range: 0.0000001% (w / v) 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), and most preferably 0.001% (w / v) to 0.2% (w / v).

[0219] In one embodiment, the pharmaceutical composition according to this disclosure may comprise 50-350 mg / mL of an antibody (e.g., an anti-HER2 antibody or an immune checkpoint antibody), a histidine buffer (providing a pH of 5.5 ± 2.0), 10-400 mM α,α-trehalose, 1-50 mM methionine, and 0.0000001% (w / v) to 0.5% (w / v) of polysorbate.

[0220] In a more specific embodiment, the pharmaceutical composition according to this disclosure may comprise 120 mg / mL of an anti-HER2 antibody or immune checkpoint antibody, 20 mM histidine buffer (providing a pH of 5.5 ± 2.0), 210 mM α,α-trehalose, 10 mM methionine, and 2,000 units / mL of the PH20 variant, and may further comprise 0.005% (w / v) to 0.1% (w / v) of polysorbate.

[0221] The pharmaceutical composition according to this disclosure can be administered by intravenous injection, subcutaneous injection, intramuscular injection, intraperitoneal injection, endothelial administration, local administration, intranasal administration, intrapulmonary administration, rectal administration, etc., and subcutaneous administration is preferably carried out by subcutaneous injection, and more preferably the pharmaceutical composition is used as an injectable preparation for subcutaneous injection.

[0222] Therefore, another embodiment of this disclosure provides a formulation comprising a pharmaceutical composition according to this disclosure, preferably an injectable formulation for subcutaneous injection.

[0223] Injectable formulations for subcutaneous injection can be provided in an immediate injection form without additional dilution and can be provided after being contained in a pre-filled syringe, glass ampoule, or plastic container.

[0224] This disclosure also relates to a method of treating a disease using a pharmaceutical composition or preparation according to this disclosure.

[0225] There are no particular limitations on the diseases that can be treated with the pharmaceutical compositions or preparations according to this disclosure, and there are no limitations on them as long as they are diseases that can be treated with a combination of a drug with a PH20 variant according to this disclosure.

[0226] The diseases that can be treated with the pharmaceutical compositions or preparations according to this disclosure may be cancer or autoimmune diseases, but are not limited thereto.

[0227] There are no particular limitations on the cancers or carcinomas that can be treated with the pharmaceutical compositions or formulations according to this disclosure, and these include solid cancers and hematologic cancers. Examples of such cancers include, but are not limited to, skin cancers (such as melanoma), liver cancer, hepatocellular carcinoma, gastric cancer, breast cancer, lung cancer, ovarian cancer, bronchial cancer, nasopharyngeal cancer, laryngeal cancer, pancreatic cancer, bladder cancer, colorectal cancer, colon cancer, cervical cancer, brain cancer, prostate cancer, bone cancer, thyroid cancer, parathyroid cancer, kidney cancer, esophageal cancer, biliary tract cancer, testicular cancer, rectal cancer, head and neck cancer, cervical cancer, ureteral cancer, osteosarcoma, neuroblastoma, fibrosarcoma, rhabdomyosarcoma, astrocytoma, neuroblastoma, and glioma. Preferably, the cancers that can be treated with the pharmaceutical compositions or formulations according to this disclosure may be selected from, but are not limited to, gastric cancer, colorectal cancer, breast cancer, lung cancer, and kidney cancer.

[0228] Autoimmune diseases that can be treated with the pharmaceutical compositions or formulations disclosed herein include, but are not limited to, rheumatoid arthritis, asthma, psoriasis, multiple sclerosis, allergic rhinitis, Crohn's disease, ulcerative colitis, systemic lupus erythematosus, type 1 diabetes, inflammatory bowel disease (IBD), and atopic dermatitis.

[0229] This disclosure also provides a method of treating a disease, comprising administering a pharmaceutical composition or preparation according to this disclosure to a subject in need of treatment, and further provides the use of the pharmaceutical composition or preparation according to this disclosure for treating a disease.

[0230] Unless otherwise defined herein, technical and scientific terms used in this disclosure have the meanings commonly understood by one of ordinary skill in the art. Furthermore, redundant descriptions of technical configurations and operations identical to those in related technologies will be omitted.

[0231] The present disclosure will be described in further detail below with reference to the following embodiments. These embodiments are provided for illustrative purposes only, and it will be clear to those skilled in the art that they should not be construed as limiting the scope of the present disclosure.

[0232] Example

[0233] Example 1. Formulation Development

[0234] As shown in Table 6, four trastuzumab subcutaneous injection formulations were prepared. Formulations 1 through 4 typically contain 120 mg / mL trastuzumab and consist of 20 mM histidine / histidine-HCl (pH 5.5), 210 mM trehalose, 10 mM methionine, and a PH20 variant. The difference between formulations 1-4 is the concentration of nonionic surfactant, where formulation 1: 0% polysorbate 20, formulation 2: 0.005% polysorbate 20, formulation 3: 0.04% polysorbate 20, and formulation 4: 0.1% polysorbate 20.

[0235] Table 6. Composition of the preparation

[0236]

[0237] Example 2. Measurement using a spectrophotometer

[0238] Preparations 1 through 4 were incubated at 45ºC for 14 days, and changes in protein concentration were analyzed using a Beckman spectrophotometer. Each sample was diluted with distilled water to a concentration of 0.4 mg / mL, and the absorbance of the protein at 280 nm was measured using the spectrophotometer. Under the harsh conditions of 14 days at 45ºC, the protein concentration of preparations 1 through 4 did not change significantly. However, hyaluronidase activity decreased rapidly at 45ºC; therefore, enzyme activity was not measured in this embodiment (see [link to documentation]). Figure 6 ).

[0239] Example 3. Size exclusion chromatography was used to study the monomer ratio of trastuzumab in each formulation.

[0240] For size exclusion chromatography analysis, an HPLC system available from Shimadzu Prominence and a TSK-gel G3000SWXL (7.8 x 300 mm, 5 μm) and a TSK guard column (6.0 x 4.0 mm, 7 μm) were used. A 0.2 M potassium phosphate solution (pH 6.2) containing 0.25 M potassium chloride was used as the mobile phase. Analysis was performed in isocratic separation mode at a flow rate of 0.5 mL / min for 35 minutes. The sample was diluted with the analytical solvent to a final concentration of 10 mg / mL, and the absorbance of the column eluent at 280 nm was recorded after injecting 20 μL into the HPLC column. The monomer ratio of trastuzumab in the HPLC chromatogram was calculated and plotted.

[0241] Size exclusion chromatography (SGC) analysis during a stability test at 45ºC for 14 days under harsh conditions showed similar patterns of change in formulations 1 through 4. The main changes were an increase in high molecular weight (HMW) and low molecular weight (LMW) degradation products and a decrease in monomer content (approximately 1.5%), with no significant differences between formulations. In summary, as a result of size exclusion chromatography analysis during a stability test at 45ºC, there were no significant differences in stability characteristics between formulations based on the concentration of polysorbate 20 (0–0.1% (w / v)) (see [link to relevant documentation]). Figure 1 ).

[0242] Example 4. Measurement of protein aggregation temperature of formulations containing trastuzumab and HP46

[0243] Dynamic light scattering (DLS) was used to analyze the denaturing properties of proteins attributed to heat. In this experiment, changes in protein molecule size with temperature variations were measured and used to calculate protein aggregation temperatures. For DLS analysis, a Zetasizer-nano-ZS instrument and quartz cuvettes (ZEN2112), available from Malvern, were used. During the analysis, the temperature was increased from 25ºC to 85ºC at 1ºC intervals, and the sample was diluted to 1 mg / mL using each prepared buffer before adding 150 µL of sample to the cuvette for analysis.

[0244] The polymerization temperature in formulation 1, which does not contain polysorbate 20, is 74ºC, and the polymerization temperature in formulations 2 to 4 is 76ºC (see [link to formulation 1]). Figure 2 ).

[0245] Example 5. WCX chromatography determination of formulations containing trastuzumab and HP46

[0246] For WCX chromatographic analysis, an HPLC system available from Shimadzu Prominence was used, employing a TSKgel CM-STAT column (4.6 x 100 mm, 7 μm) and a TSKgel protective gel CMSTAT (3.2 mm inner diameter x 1.5 cm). Mobile phase A was 10 mM sodium phosphate (pH 7.5), and mobile phase B was 10 mM sodium phosphate containing 0.1 M NaCl (pH 7.2). Analysis was performed at a flow rate of 0.8 mL / min for 55 minutes along a linear concentration gradient of 0–30% mobile phase B. The sample was diluted with mobile phase A to a final concentration of 1.0 mg / mL, and 80 μL of the sample was injected into the HPLC system. The absorbance of the column eluent at 280 nm was then recorded. The monomer ratio of trastuzumab in the HPLC chromatogram was calculated and plotted.

[0247] When WCX analysis was performed under harsh conditions (45ºC) for 14 days, formulations 1 through 4 showed similar patterns of change. Specific changes included an increase in the relative content of the acidic variant (approximately 30% change after 14 days), a decrease in the relative content of the main peak (approximately 44% change after 14 days), and an increase in the relative content of the basic variant (approximately 15% change after 14 days), with no significant differences observed across formulations. In summary, protein stability under harsh conditions (45ºC) for WCX analysis of polysorbate 20 (0–0.1%) is similar (see [link to relevant documentation]). Figure 3 ).

[0248] Example 6. Formulation Development

[0249] As shown in Table 7, three types of trastuzumab subcutaneous injection formulations were prepared. Formulations 5 through 7 typically contain 120 mg / mL trastuzumab, 20 mM histidine / histidine-HCl (pH 5.5), 210 mM trehalose, 10 mM methionine, and HP46. The difference between formulations 5-7 is the composition of stabilizer 3: formulation 5: 0.04% polysorbate 20, formulation 6: 50 mmLys-Lys, and formulation 3: glycine.

[0250] Table 7. Composition of the preparation

[0251]

[0252] Example 7. Measurement using a spectrophotometer

[0253] Preparations 5 to 7 were incubated at 45ºC for 14 days, and changes in protein concentration were analyzed using a Beckman spectrophotometer. Each sample was diluted with distilled water to a concentration of 0.4 mg / mL, and the absorbance of the protein at 280 nm was measured using a spectrophotometer. Under the harsh conditions of 14 days at 45ºC, the protein concentration of preparations 5 to 7 did not change significantly. However, hyaluronidase activity decreased rapidly at 45ºC; therefore, enzyme activity was not measured in this embodiment (see [link to documentation]). Figure 6 ).

[0254] Example 8. The monomer ratio of trastuzumab in each formulation was studied using size exclusion chromatography.

[0255] For size exclusion chromatography analysis, an HPLC system available from Shimadzu Prominence was used, with a TSK-gel G3000SWXL (7.8 x 300 mm, 5 μm) and a TSK guard column (6.0 x 4.0 mm, 7 μm) as the column. A 0.2 M potassium phosphate solution (pH 6.2) containing 0.25 M potassium chloride was used as the mobile phase. Isocratic separation mode was applied at a flow rate of 0.5 mL / min for 35 minutes. The sample was diluted with the analytical solvent to a final concentration of 10 mg / mL, and the absorbance at 280 nm was measured after injecting 20 μL of sample into the HPLC column. The monomer ratio of trastuzumab in the HPLC chromatogram was calculated and plotted.

[0256] Size exclusion chromatography (SGC) analysis of formulations 5 through 7 showed similar patterns of change during a stability test conducted under harsh conditions (45°C) for 14 days. The main changes were an increase in high molecular weight (HMW) and low molecular weight (LMW) impurities and a decrease in monomer content (approximately 1.5%), and these differences were not significant depending on the formulation. In summary, as results of size exclusion chromatography analysis during stability testing under harsh conditions (i.e., 45°C), formulations of 0.04% polysorbate 20, 50 mM Lys-Lys, and 50 mM glycine showed similar protein stability (see [link to product description]). Figure 4 ).

[0257] Example 9. WCX chromatographic analysis of formulations containing trastuzumab and HP46

[0258] For WCX chromatographic analysis, an HPLC system available from Shimadzu Prominence was used, with TSKgel CM-STAT (4.6 × 100 mm, 7 μm) and TSKgel protective gel CMSTAT (3.2 mm inner diameter x 1.5 cm) as columns. Mobile phase A was 10 mM sodium phosphate (pH 7.5), and mobile phase B was 10 mM sodium phosphate containing 0.1 M NaCl (pH 7.2). Analysis was performed for 55 minutes using a 0-30% linear concentration gradient at a flow rate of 0.8 mL / min. The sample was diluted with mobile phase A to a final concentration of 1.0 mg / mL, and 80 μL of the sample was injected into the HPLC system. The absorbance was then recorded at 280 nm. The monomer ratio of trastuzumab in the HPLC chromatogram was calculated and plotted.

[0259] When WCX analysis was performed under harsh conditions (i.e., 45ºC) for 14 days, formulations 5 through 7 showed similar patterns of change. Specific changes included an increase in the relative content of the acidic variant (approximately 30% change over 14 days), a decrease in the relative content of the main peak (approximately 44% change over 14 days), and an increase in the relative content of the basic variant (approximately 15% change over 14 days), with no significant differences depending on the formulation. In summary, as a result of WCX analysis under harsh conditions (i.e., 45ºC), formulations of 0.04% polysorbate 20, 50 mM Lys-Lys, and 50 mM glycine showed similar protein stability (see [link to relevant documentation]). Figure 5 ).

[0260] Example 10. Stability of HP46 was evaluated at 40ºC and 45ºC in subcutaneous injection formulations of trastuzumab and HP46.

[0261] To assess the stability of HP46 in the trastuzumab subcutaneous injection formulation, trastuzumab (120 mg / mL) and pH20 (2000 units / mL) were mixed. The buffer used contained 20 mM histidine (pH 5.5), 210 mM trehalose, 10 mM methionine, and 0.04% polysorbate 20. Enzyme activity in control samples was measured on day 0, and experimental samples were incubated at 40ºC or 45ºC for 1 day, after which enzyme activity was measured for each sample.

[0262] Herceptin subcutaneous injection formulations of trastuzumab + HW2 and trastuzumab + HP46 were each incubated at 40ºC for 1 day, and hyaluronidase activity was then measured. The results showed 51%, 47%, and 94% activity in each case, respectively, indicating that HP46 exhibits the greatest thermostability at 40ºC (see [link to product description]). Figure 6 Additionally, the Herceptin subcutaneous formulations trastuzumab + HW2 and trastuzumab + HP46 were incubated at 45ºC for 1 day, and hyaluronidase activity was measured. As a result, the Herceptin subcutaneous formulation and trastuzumab + HW2 showed no enzyme activity, but trastuzumab + HP46 retained its enzyme activity (see [link to original text]). Figure 6 ).

[0263] Example 11. Formulation Development

[0264] As shown in Table 8, three trastuzumab subcutaneous injection formulations were prepared. Formulations 8 to 10 typically contain 120 mg / mL trastuzumab, 20 mM histidine / histidine-HCl (pH 5.5), 210 mM trehalose, 10 mM methionine, and a pH20 variant. The difference between formulations 8 and 10 is the concentration of nonionic surfactants: formulation 8: 0% polysorbate 20, formulation 9: 0.005% polysorbate 20, and formulation 10: 0.04% polysorbate 20.

[0265] Table 8. Composition of the preparation

[0266]

[0267] Example 12. Measurement using a spectrophotometer

[0268] Preparations 8 to 10 were incubated at 40ºC for 14 days, and changes in protein concentration were analyzed using a Beckman spectrophotometer. Each sample was diluted with distilled water to a concentration of 0.4 mg / mL, and the absorbance of the protein at 280 nm was measured using a spectrophotometer. Under the harsh conditions of 14 days at 40ºC, the protein concentration of preparations 8 to 10 showed no significant change.

[0269] Example 13. The monomer ratio of trastuzumab in each formulation was studied using size exclusion chromatography.

[0270] For size exclusion chromatography analysis, an HPLC system available from Shimadzu Prominence was used, with a TSK-gel G3000SWXL (7.8 x 300 mm, 5 μm) and a TSK guard column (6.0 x 4.0 mm, 7 μm) as the column. A 0.2 M potassium phosphate (pH 6.2) containing 0.25 M potassium chloride was used as the mobile phase. Analysis was performed in isocratic separation mode at a flow rate of 0.5 mL / min for 35 minutes. The sample was diluted with the analytical solvent to a final concentration of 10 mg / mL, and the absorbance at 280 nm was measured after injecting 20 μL of sample into the HPLC column. The monomer ratio of trastuzumab in the HPLC chromatogram was calculated and plotted.

[0271] Size exclusion chromatography (SGC) analysis during a stability test at harsh conditions (40ºC) for 14 days showed similar patterns of change in formulations 8 through 10. The main changes were an increase in high molecular weight (HMW) and low molecular weight (LMW) degradation products and a decrease in monomer content (approximately less than 1.0%), and no significant differences were observed between formulations. In summary, as a result of size exclusion chromatography analysis during a stability test at harsh conditions (40ºC), there were no significant differences in the overall stability profile between formulations based on the concentration of polysorbate 20 (0–0.04%) (see [link to relevant documentation]). Figure 7 ).

[0272] Example 14. Measurement of protein aggregation temperature in formulations containing trastuzumab and HP46

[0273] Dynamic light scattering (DLS) was used to analyze the denaturation properties of proteins attributed to heat in the field of protein pharmaceuticals. In this experiment, changes in protein molecule size with temperature variations were measured and used to calculate the protein aggregation temperature. For DLS analysis, a Zetasizer-nano-ZS instrument and quartz cuvettes (ZEN2112), available from Malvern, were used. During the analysis, the temperature was increased from 25ºC to 85ºC at 1ºC intervals, and the sample was diluted to 1 mg / mL using each preparation buffer before adding 150 µL of sample to the cuvette for analysis.

[0274] The aggregation temperature of formulation 8 (without polysorbate 20) was 78.3ºC, that of formulation 9 was 77.3ºC, and that of formulation 10 was 77.7ºC. In Example 13, despite the absence of polysorbate 20, the monomer ratio of the protein did not show any change, and as a comparison between the cases without and with polysorbate 20, no difference in protein aggregation was confirmed. These results indicate that subcutaneous injection formulations of trastuzumab do not necessarily require a minimum amount of polysorbate 20 (see [link to example]). Figure 8 ).

[0275] Example 15. WCX chromatographic analysis of formulations containing trastuzumab and HP46.

[0276] For WCX chromatographic analysis, an HPLC system available from Shimadzu Prominence was used, employing a TSKgel CM-STAT column (4.6 x 100 mm, 7 μm) and a TSKgel protective gel CMSTAT (3.2 mm inner diameter x 1.5 cm). Mobile phase A was 10 mM sodium phosphate (pH 7.5), and mobile phase B was 10 mM sodium phosphate containing 0.1 M NaCl (pH 7.2). Analysis was performed at a flow rate of 0.8 mL / min for 55 minutes along a linear concentration gradient of 0–30% mobile phase B. The sample was diluted with mobile phase A to a final concentration of 1.0 mg / mL, and 80 μL of the sample was injected into the HPLC system. The absorbance of the column eluent at 280 nm was then recorded. The monomer ratio of trastuzumab in the HPLC chromatogram was calculated and plotted.

[0277] When WCX analysis was performed under harsh conditions (40ºC) for 14 days, formulations 8 through 10 showed similar patterns of change. Specific changes included an increase in the relative content of the acidic variant (approximately 10% change over 14 days), a decrease in the relative content of the main peak (approximately 40% change over 14 days), and an increase in the relative content of the basic variant (approximately 300% change over 14 days), with no significant differences observed across formulations. In summary, the protein stability of polysorbate 20 (0–0.04%) was similar under harsh conditions (40ºC) in WCX analysis (see Figure 9).

[0278] Example 16. Enzyme activity measurement of formulations containing trastuzumab and HP46

[0279] The turbidity assay used to measure enzyme activity is a method that measures the degree of aggregate formation by binding residual hyaluronic acid to acidified albumin (BSA) via absorbance. When hyaluronic acid is hydrolyzed by pH 20, the degree of binding to albumin decreases, resulting in a decrease in absorbance. BTH (Sigma), as a standardized product, was diluted to 1, 2, 5, 7.5, 10, 15, 20, 30, 50, and 60 units / mL and prepared in each tube. Purified pH 20 variant samples were diluted with enzyme dilution buffer (20 mM Tris·HCl pH 7.0, 77 mM NaCl, 0.01% (w / v) bovine serum albumin) to 100X, 300X, 600X, 1200X, and 2400X and prepared in each tube. In fresh test tubes, dilute the 3 mg / mL hyaluronidase solution 10-fold to a concentration of 0.3 mg / mL, making each tube 180 µL. Add 60 μL of the sample containing hyaluronidase to the diluted hyaluronic acid solution and mix, allowing the reaction to proceed at 37ºC for 45 minutes. After the reaction is complete, add 50 μL of the reacted enzyme and 250 μL of acidic albumin solution to each well of a 96-well plate and shake for 10 minutes. Then, measure the absorbance at 600 nm using a spectrophotometer.

[0280] The results of activity analysis, conducted under harsh conditions (40ºC for 14 days), confirmed that higher concentrations of polysorbate 20 correlated with a greater decrease in activity over time (see [link to relevant documentation]). Figure 10 ).

[0281] Example 17. Formulation Development

[0282] As shown in Table 9, three trastuzumab subcutaneous injection formulations were prepared. Formulations 11 to 13 typically contain 120 mg / mL trastuzumab, 20 mM histidine / histidine-HCl (pH 5.5), 210 mM trehalose, 10 mM methionine, and a PH20 variant. The difference between formulations 11-13 is the concentration of nonionic surfactant, with formulation 11 containing 0% polysorbate 80, formulation 12 containing 0.005% polysorbate 80, and formulation 13 containing 0.04% polysorbate 80.

[0283] Table 9. Composition of the preparation

[0284]

[0285] Size exclusion chromatography (SGC) analysis during a stability test at harsh conditions (40ºC) for 14 days showed similar patterns of change in formulations 11 through 13. The main changes were an increase in high molecular weight (HMW) and low molecular weight (LMW) degradation products and a decrease in monomer content (approximately less than 1.0%), and no significant differences were observed between formulations. In summary, as a result of size exclusion chromatography analysis during a stability test at harsh conditions (40ºC), there were no significant differences in stability characteristics between formulations based on the concentration of polysorbate 80 (0–0.04%) (see [link to relevant documentation]). Figure 11 ).

[0286] Example 18. WCX chromatographic analysis of a formulation containing trastuzumab and HP46.

[0287] For WCX chromatographic analysis, an HPLC system available from Shimadzu Prominence was used, employing a TSKgel CM-STAT column (4.6 x 100 mm, 7 μm) and a TSKgel protective gel CMSTAT (3.2 mm inner diameter x 1.5 cm). Mobile phase A was 10 mM sodium phosphate (pH 7.5), and mobile phase B was 10 mM sodium phosphate containing 0.1 M NaCl (pH 7.2). Analysis was performed at a flow rate of 0.8 mL / min for 55 minutes along a linear concentration gradient of 0–30% mobile phase B. The sample was diluted with mobile phase A to a final concentration of 1.0 mg / mL, and 80 μL of the sample was injected into the HPLC system. The absorbance of the column eluent at 280 nm was then recorded. The monomer ratio of trastuzumab in the HPLC chromatogram was calculated and plotted.

[0288] When WCX analysis was performed under harsh conditions, i.e., a 14-day stability test at 40ºC, formulations 11 through 13 showed similar patterns of change. Specific changes included an increase in the relative content of the acidic variant (approximately 10% change over 14 days), a decrease in the relative content of the main peak (approximately 40% change over 14 days), and an increase in the relative content of the basic variant (approximately 300% change over 14 days), with no significant differences observed across formulations. In summary, the protein stability of polysorbate 80 (0–0.04%) was similar under harsh conditions, i.e., a stability test at 40ºC, according to WCX analysis (see Figure 12).

[0289] Example 19. Enzyme activity measurement of formulations containing trastuzumab and HP46

[0290] The turbidity assay used to measure enzyme activity is a method that measures the degree of aggregation of residual hyaluronic acid bound to acidified albumin (BSA) by absorbance. When hyaluronic acid is hydrolyzed at pH 20, the degree of binding to albumin decreases, resulting in a decrease in absorbance. BTH (Sigma), as a standardized product, was diluted to 1, 2, 5, 7.5, 10, 15, 20, 30, 50, and 60 units / mL, and prepared in each tube. Purified protein samples were diluted with enzyme dilution buffer (20 mM Tris·HCl pH 7.0, 77 mM NaCl, 0.01% (w / v) bovine serum albumin) to 100X, 300X, 600X, 1200X, and 2400X, and prepared in each tube. In fresh test tubes, dilute the 3 mg / mL hyaluronidase solution 10-fold to a concentration of 0.3 mg / mL, making each tube 180 µL. Add 60 μL of the sample containing hyaluronidase to the diluted hyaluronic acid solution, mix, and allow to react at 37ºC for 45 minutes. After the reaction is complete, add 50 μL of the reacted enzyme and 250 μL of acidic albumin solution to each well of a 96-well plate and shake for 10 minutes. Then, measure the absorbance at 600 nm using a spectrophotometer.

[0291] The results of activity analysis, conducted under harsh conditions (40ºC for 14 days), confirmed that higher concentrations of polysorbate 80 correlated with a greater decrease in activity over time (see [link]). Figure 13 ).

[0292] Example 20. Formulation Development

[0293] As described in Table 10, three types of rituximab formulations were prepared. Formulations 14 to 16 typically contain 120 mg / mL rituximab, 20 mM histidine / histidine-HCl (pH 5.5), 210 mM trehalose, 10 mM methionine, and a PH20 variant. The difference between formulations 14-16 is the concentration of the nonionic surfactant: formulation 14: 0% polysorbate 80, formulation 15: 0.005% polysorbate 80, and formulation 16: 0.06% polysorbate 80.

[0294] Table 10. Composition of the preparation

[0295]

[0296] Size exclusion chromatography (SGC) analysis during a stability test at 40ºC for 7 days under harsh conditions showed similar patterns of change in formulations 14 through 16. The main changes were an increase in high molecular weight (HMW) and low molecular weight (LMW) degradation products and a decrease in monomer content (less than approximately 1.0%), with no significant differences between formulations. In summary, as a result of size exclusion chromatography analysis during a stability test at 40ºC, there were no significant differences in the overall stability profile between formulations based on the concentration of polysorbate 80 (0–0.06%) (see [link to relevant documentation]). Figure 14 ).

[0297] Example 21. Measurement of enzyme activity in formulations containing rituximab and HP46

[0298] The turbidity assay used to measure enzyme activity is a method that measures the degree of aggregation of residual hyaluronic acid bound to acidified albumin (BSA) by absorbance. When hyaluronic acid is hydrolyzed at pH 20, the degree of binding to albumin decreases, resulting in a decrease in absorbance. BTH (Sigma), as a standardized product, was diluted to 1, 2, 5, 7.5, 10, 15, 20, 30, 50, and 60 units / mL, and prepared in each tube. Purified protein samples were diluted with enzyme dilution buffer (20 mM Tris·HCl pH 7.0, 77 mM NaCl, 0.01% (w / v) bovine serum albumin) to 100X, 300X, 600X, 1200X, and 2400X, and prepared in each tube. In fresh test tubes, dilute the 3 mg / mL hyaluronidase solution 10-fold to a concentration of 0.3 mg / mL, making each tube 180 µL. Add 60 μL of the sample containing hyaluronidase to the diluted hyaluronic acid solution, mix, and allow to react at 37ºC for 45 minutes. After the reaction is complete, add 50 μL of the reacted enzyme and 250 μL of acidic albumin solution to each well of a 96-well plate and shake for 10 minutes. Then, measure the absorbance at 600 nm using a spectrophotometer.

[0299] The results of activity analysis, conducted under harsh conditions (7 days at 40ºC), confirmed that higher concentrations of polysorbate 80 correlated with a greater decrease in activity over time (see [link to relevant documentation]). Figure 15 ).

[0300] Example 22. Measurement of enzyme activity in formulations containing commercially available polysorbate-free products.

[0301] As described in Table 11, two types of commercially available rituximab formulations were prepared. Formulation 17 is a commercially available buffer for subcutaneous injection of the formulation, and formulation 18 is a commercially available buffer for intravenous injection of the formulation. Formulations 17 and 18 contain 120 mg / mL of the PH20 variant and 100 mg / mL of rituximab, respectively, but unlike the commercially available formulations, they do not contain polysorbate 80.

[0302] Table 11. Composition of the preparation

[0303]

[0304] The turbidity assay used to measure enzyme activity is a method that measures the degree of aggregation of residual hyaluronic acid bound to acidified albumin (BSA) by absorbance. When hyaluronic acid is hydrolyzed at pH 20, the degree of binding to albumin decreases, resulting in a decrease in absorbance. BTH (Sigma), as a standardized product, was diluted to 1, 2, 5, 7.5, 10, 15, 20, 30, 50, and 60 units / mL, and prepared in each tube. Purified protein samples were diluted with enzyme dilution buffer (20 mM Tris·HCl pH 7.0, 77 mM NaCl, 0.01% (w / v) bovine serum albumin) to 100X, 300X, 600X, 1200X, and 2400X, and prepared in each tube. In fresh test tubes, dilute the 3 mg / mL hyaluronidase solution 10-fold to a concentration of 0.3 mg / mL, making each tube 180 µL. Add 60 μL of the sample containing hyaluronidase to the diluted hyaluronic acid solution, mix, and allow to react at 37ºC for 45 minutes. After the reaction is complete, add 50 μL of the reacted enzyme and 250 μL of acidic albumin solution to each well of a 96-well plate and shake for 10 minutes. Then, measure the absorbance at 600 nm using a spectrophotometer.

[0305] As a result of activity analysis conducted under harsh conditions (i.e., at 40ºC) for 6 consecutive days, it was confirmed that high activity was maintained even in formulations without polysorbate 80, and in particular, formulation 18 maintained high activity (see [link to relevant documentation]). Figure 16 ).

[0306] Example 23: Formulation Development

[0307] As described in Table 12, four types of pembrolizumab formulations were prepared. Formulations 19, 20, and 21 typically contain 25 mg / mL pembrolizumab, 10 mM histidine (pH 5.5), 7% sucrose, 10 mM methionine, and the PH20 variant. The difference between formulations 19-21 is the concentration of nonionic surfactant: formulation 19: 0% polysorbate 80, formulation 20: 0.005% polysorbate 80, and formulation 21: 0.02% polysorbate 80. Formulation 22 contains 25 mg / mL pembrolizumab and consists of 10 mM histidine (pH 5.5), 210 mM trehalose, 10 mM methionine, 0.02% polysorbate 80, and the PH20 variant.

[0308] Table 12. Composition of the preparation

[0309]

[0310] Example 24. Measurement using a spectrophotometer

[0311] Preparations 19, 20, 21, and 22 were incubated at 40ºC for 7 days, and changes in protein concentration were analyzed using a Beckman spectrophotometer. Each sample was diluted with distilled water to a concentration of 0.4 mg / mL, and the absorbance of the protein at 280 nm was measured using a spectrophotometer.

[0312] In a stability test conducted under harsh conditions, namely at 40ºC for 7 consecutive days, the protein concentration of formulations 19 to 22 did not change significantly.

[0313] Example 25. Size exclusion chromatography was used to study the monomer ratio of pembrolizumab in each formulation.

[0314] For size exclusion chromatography analysis, an HPLC system available from Shimadzu Prominence was used, with a TSK-gel G3000SWXL (7.8 x 300 mm, 5 μm) and a TSK guard column (6.0 x 4.0 mm, 7 μm) as the column. A 0.2 M potassium phosphate (pH 6.2) containing 0.25 M potassium chloride was used as the mobile phase. Analysis was performed in isocratic separation mode at a flow rate of 0.5 mL / min for 35 minutes. The sample was diluted with the analytical solvent to a final concentration of 10 mg / mL, and the absorbance of the column eluent at 280 nm was measured after injecting 20 μL of sample into the HPLC column. The monomer ratio of pembrolizumab in the HPLC chromatogram was calculated and plotted.

[0315] Size exclusion chromatography (SGC) analysis of formulations under harsh conditions (40ºC) for 7 days showed similar patterns of change in formulations 19, 20, 21, and 22. There were no significant differences in the patterns of change for high molecular weight (HMW) and low molecular weight (LMW) degradation products, depending on the formulation. In summary, as results of size exclusion chromatography analysis under harsh conditions (40ºC), formulations 19, 20, 21, and 22 showed no significant differences, and there were no differences based on sugar type (see [link to relevant documentation]). Figure 17 These results are consistent with those of trastuzumab and rituximab according to previous embodiments.

[0316] Example 26. Measurement of enzyme activity in formulations containing pembrolizumab and HP46

[0317] The turbidity assay used to measure enzyme activity is a method that measures the degree of aggregation of residual hyaluronic acid bound to acidified albumin (BSA) by absorbance. When hyaluronic acid is hydrolyzed at pH 20, the degree of binding to albumin decreases, resulting in a decrease in absorbance. BTH (Sigma), as a standardized product, was diluted to 1, 2, 5, 7.5, 10, 15, 20, 30, 50, and 60 units / mL, and prepared in each tube. Purified protein samples were diluted with enzyme dilution buffer (20 mM Tris·HCl pH 7.0, 77 mM NaCl, 0.01% (w / v) bovine serum albumin) to 100X, 300X, 600X, 1200X, and 2400X, and prepared in each tube. In fresh test tubes, dilute the 3 mg / mL hyaluronidase solution 10-fold to a concentration of 0.3 mg / mL, making each tube 180 µL. Add 60 μL of the enzyme-containing sample to the diluted hyaluronic acid solution, mix, and allow to react at 37ºC for 45 minutes. After the reaction is complete, add 50 μL of the reacted enzyme and 250 μL of acidic albumin solution to each well of a 96-well plate and shake for 10 minutes. Then, measure the absorbance at 600 nm using a spectrophotometer.

[0318] The results of activity analysis, conducted under harsh conditions (7 days at 40ºC), confirmed that the decrease in activity over time increased with increasing polysorbate 80 concentration. It was also confirmed that, when containing the same amount of polysorbate 80, the decrease in activity in trehalose-containing formulations was less than that in sucrose-containing formulations (see [link to product description]). Figure 18 ).

[0319] Example 27. pH-activity profiles of HP46 and wild-type HW2

[0320] For experiments conducted to determine the pH-activity profiles of HP46 and wild-type HW2, microturbidity assays were used. For each pH, ​​hyaluronic acid buffer was prepared to dissolve hyaluronic acid as the substrate, and enzyme buffer was prepared to dilute the enzyme.

[0321] Three 96-well plates were prepared for the enzyme-substrate reaction and named A, B, and C, and experiments were conducted.

[0322] Hyaluronic acid buffers with pH values ​​of 4.0, 4.5, or 5.0 were prepared using 20 mM acetic acid and 70 mM NaCl, and hyaluronic acid solutions with pH values ​​of 5.5, 6.0, 6.5, 7.0, or 8.0 were prepared using 20 mM sodium phosphate and 70 mM NaCl. 20 mg of hyaluronic acid was dissolved in 10 mL of each of the prepared hyaluronic acid buffers to prepare the final hyaluronic acid substrate solution. This solution was then diluted with the prepared hyaluronic acid buffers according to the pH values ​​to concentrations of 0.1 mg / mL, 0.25 mg / mL, 0.45 mg / mL, or 0.7 mg / mL to prepare 500 µL solutions. 100 µL of each solution was aliquoted into each well of a 96-well plate designated A. The concentration-diluted and prepared hyaluronic acid buffers were used as calibration curves for measuring hyaluronic acid concentrations.

[0323] Enzyme buffers with pH 4.0, 4.5, or 5.0 were prepared using 20 mM acetic acid, 0.01% (w / v) BSA, and 70 mM NaCl, and enzyme buffers with pH 5.5, 6.0, 6.5, 7.0, or 8.0 were prepared using 20 mM sodium phosphate, 0.01% (w / v) BSA, and 70 mM NaCl.

[0324] HP46 and wild-type HW2 enzymes were diluted to 10 units / mL with an enzyme buffer prepared according to pH, and then 50 µL of the resulting solution was dispensed into each well of a 96-well plate named B.

[0325] 50 µL of sample was transferred from each well of a 96-well plate named A to each well of a 96-well plate named B, and then allowed to react in a 37ºC shaking incubator for 45 minutes. 15 minutes before the reaction was complete, 200 µL of acidic albumin solution was dispensed into each well of a 96-well plate named C for preparation, and when the enzyme substrate reaction was complete, 40 µL of sample was transferred from each well of the 96-well plate named B to each well of the 96-well plate named C, and then allowed to react for 20 minutes. After 20 minutes, absorbance was measured at 600 nm, and the amount of residual hyaluronic acid after the enzyme substrate reaction was calculated. Enzyme activity curves based on pH were also constructed (see [link to relevant documentation]). Figure 19 ).

[0326] Example 28. Pharmacokinetic studies of Herceptin subcutaneous formulation, trastuzumab, and HP46 in Sprague-Dawley rats.

[0327] To examine whether the subcutaneous formulations of trastuzumab and HP46 exhibited the same pharmacokinetic properties as the subcutaneous formulation of Herceptin, 9-week-old Sprague-Dawley rats were used. Herceptin and trastuzumab were administered at a dose of 18 mg / kg rat body weight. The subcutaneous formulation of Herceptin contained 100 U of rHuPH20, and the amount of HP46 was also 100 U. In pharmacokinetic tests, trastuzumab and HP46 showed the same area under the curve (AUC) as the subcutaneous formulation of Herceptin (see [link to pharmacokinetic study]). Figure 20 ).

[0328] [Industrial Applicability]

[0329] The pharmaceutical composition according to this disclosure can be used for subcutaneous injection and is very stable, and the activity of the PH20 variant, along with the drug, preferably an antibody drug, can be maintained for a long time. Therefore, the pharmaceutical composition can not only help reduce the production cost of subcutaneous injection preparations, but also reduce medical costs, and is very effective in terms of patient convenience.

[0330] Although preferred embodiments of this disclosure have been disclosed for illustrative purposes, those skilled in the art will understand that various modifications, additions, and substitutions may be made without departing from the scope and spirit of the invention as disclosed in the appended claims.

[0331] References

[0332] 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 transportof therapeutics. J Control Release 114, 230-241.

[0333] Borders jr., C.L. and Raftery, A. (1968) Purification and PartialCharacterization of Testicular Hyaluronidase. J Biol Chem 243, 3756-3762

[0334] Chao, K.L., Muthukumar, L., and Herzberg, O. (2007). Structure ofhuman hyaluronidase-1, a hyaluronan hydrolyzing enzyme involved in tumorgrowth and angiogenesis. Biochemistry 46, 6911-6920.

[0335] Chen, K.J., Sabrina, S., El-Safory, N.S., Lee, G.C., and Lee, C.K.(2016) Constitutive expression of recombinant human hyaluronidase PH20 byPichia pastoris. J Biosci Bioeng. 122, 673-678

[0336] Frost, GI (2007). Recombinant human hyaluronidase (rHuPH20): anenabling platform for subcutaneous drug and fluid administration. Expert OpinDrug Deliv 4, 427-440

[0337] Hofinger, ES, Bernhardt, G., and Buschauer, A. (2007) Kinetics ofHyal-1 and PH-20 hyaluronidases: comparison of minimal substrates and analysis of the transglycosylation reaction. Glycobiology 17, 963-971

[0338] Kreidieh, FY, Moukadem, HA, and Saghir, NSE (2016) Overview, prevention and management of chemotherapy extravasation. World J Clin Oncol7, 87-97.

[0339] Thomas, JR, Yocum, RC, Haller, MF, and Flament J. (2009) TheINFUSE-Morphine IIB Study: Use of Recombinant Human Hyaluronidase (rHuPH20) to Enhance the Absorption of Subcutaneous Morphine in Healthy Volunteers. JPain Symptom Manag 38, 673-682

[0340] text file

[0341] See the attached sequence list <110> Atgen Company <120> A pharmaceutical composition comprising a human hyaluronidase PH20 variant and a 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 Ala 1 5 10 15 Tyr Ser <210> 4 <211> twenty one <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 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 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>[[ID=2,8]] [[ID=,2,9]]<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 Note: There seems to be a formatting issue with the original text where tags might be overlapping in a non-standard way. I've tried to translate it as accurately as possible while maintaining the integrity of the tags. If this is a specific format for a particular system, it might be beneficial to double-check the original source for proper formatting guidelines. 65 70 75 80 Gln Lys Ile Ser Leu Gln Asp His Leu Asp Lys Ala 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 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 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 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 ​​​​​​​​​​​​​​​​​​​​​ 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 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 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 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 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<00~01470><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 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 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 [[ID=2�]] 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 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 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 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[[ID=is=18]] 450 <210> 21 <211> 452 <212> PRT <213> Artificial Sequence <220> <223> Synthetic Sequence ​​​​​​​​​​​​​​​​​​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<000)2041><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 请注意,原文中`<000)2041>`这里括号可能是多余的,已按原样翻译。你可再核对下原文准确性。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 sixty 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 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 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 Pro1] 65 70 75 80 Gln Lys Ile Ser Leu Gln Asp His Leu Asp Lys Ala 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 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 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 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 谷氨酸 酪氨酸 甲硫氨酸 天冬氨酸 苏氨酸 苏氨酸 亮氨酸 天冬酰胺 脯氨酸 酪氨酸 异亮氨酸 异亮氨酸 天冬酰胺 缬氨酸 苏氨酸 亮氨酸 325 330 335 丙氨酸 丙氨酸 赖氨酸 甲硫氨酸 半胱氨酸 丝氨酸 谷氨酰胺 缬氨酸 亮氨酸 半胱氨酸 谷氨酰胺 谷氨酸 谷氨酰胺 甘氨酸 缬氨酸 半胱氨酸 340 345 350 异亮氨酸 精氨酸 赖氨酸 天冬酰胺 色氨酸 天冬酰胺 丝氨酸 丝氨酸 天冬氨酸 酪氨酸 亮氨酸 组氨酸 亮氨酸 天冬酰胺 脯氨酸 天冬氨酸 355 360 365 天冬酰胺 苯丙氨酸 丙氨酸 异亮氨酸 谷氨酰胺 亮氨酸 谷氨酸 赖氨酸 甘氨酸 甘氨酸 赖氨酸 苯丙氨酸 苏氨酸 缬氨酸 精氨酸 甘氨酸 370 375 380 赖氨酸 脯氨酸 苏氨酸 亮氨酸 谷氨酸 天冬氨酸 亮氨酸 谷氨酸 谷氨酰胺 苯丙氨酸 丝氨酸 谷氨酸 赖氨酸 苯丙氨酸 酪氨酸 半胱氨酸 385 390 395 400 丝氨酸 半胱氨酸 酪氨酸 丝氨酸 苏氨酸 亮氨酸 丝氨酸 半胱氨酸 赖氨酸 谷氨酸 赖氨酸 丙氨酸 天冬氨酸 缬氨酸 赖氨酸 天冬氨酸 405 410 415 苏氨酸 天冬氨酸 丙氨酸 缬氨酸 420 <210> 32 <211> 433 <212> PRT <213> Artificial Sequence <220> <223> Synthetic Sequence <400> 32 苯丙氨酸 精氨酸 丙氨酸 脯氨酸 脯氨酸 缬氨酸 异亮氨酸 脯氨酸 天冬酰胺 缬氨酸 脯氨酸 苯丙氨酸 亮氨酸 色氨酸 丙氨酸 色氨酸 1 5 10 15 天冬酰胺 丙氨酸 脯氨酸 丝氨酸 谷氨酸 苯丙氨酸 半胱氨酸 亮氨酸 甘氨酸 赖氨酸 苯丙氨酸 天冬氨酸 谷氨酸 脯氨酸 亮氨酸 天冬氨酸 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> Synthetic 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> Synthetic 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> Synthetic 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> [[ID=​​​​​​​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 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

Claims

1. A pharmaceutical composition comprising: (a) medicines; and (b) a PH20 variant at a concentration of 100 to 20,000 units / mL, wherein the PH20 variant consists of the amino acid sequence of SEQ ID NO:

44.

2. The pharmaceutical composition according to claim 1, wherein the drug is a protein drug, an antibody, a small molecule, an aptamer, RNAi, an antisense or a cell therapy agent.

3. The pharmaceutical composition according to claim 1, wherein the drug is an antibody. The pharmaceutical composition according to claim 1 , wherein the drug is a soluble receptor or an Fc fusion protein thereof.

5. The pharmaceutical composition according to claim 3, wherein the antibody is anti-CTLA4, anti-HER2, anti-PD-1, anti-PD-L1, anti-TNFα or anti-CD20 antibody.

6. The pharmaceutical composition of claim 3, wherein the antibody binds to one or more antigens selected from the group consisting of 4-1BB, integrin, beta-amyloid, angiopoietin, angiopoietin analog 3, 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, tight junction protein-18, complement factor D, CTLA4, DLL3, EGF receptor, hemophilia factor, F c 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 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 vWF.

7. The pharmaceutical composition according to claim 3, wherein the antibody is an immune checkpoint antibody.

8. The pharmaceutical composition of claim 3, wherein the antibody is trastuzumab, pertuzumab, rituximab, infliximab, adalimumab, golimumab, ipilimumab, nivolumab, avelumab, atezolizumab, or durvalumab.

9. The pharmaceutical composition according to claim 1, wherein the drug is etanercept.

10. The pharmaceutical composition according to claim 3, wherein the antibody is one or more selected from the group consisting of: utoluzumab, natalizumab, etrolizumab, vedolizumab, biglumab, bapiluzumab, crenetuzumab, sorazumab, aducanumab, gantezumab, AMG 780, MEDI 3617, neskamab, vancouverizumab, ivacitumomab, tamifluzumab, lanarumab, belimumab, obinutuzumab, ravulizumab, eculizumab, molizumab, oxicizumab, tilizumab, muromonab, tebentafusp, blinatumomab, REGN1979, ibalizumab, zalimumab, itrazumab, efalizumab, inbilizumab, tafasitumomab, teloximab, ocrelizumab, utiluximab, obinutuzumab, ofatumumab, rituximab, tositumomab, ibritumomab tiuxetan, epratuzumab, inotuzumab, pasituzumab, bentuozumab tadalafil-vadatuximab, ozogamicin-gemtuzumab, daratumumab, isatuximab, alemtuzumab, rizanizumab, pollotuzumab, galiximab, eprenazumab, frenazumab, gacapatuzumab, erenomab, zoloftizumab, lambazumab, tremelimumab, zeflalimumab, ipilimumab, rovaceptuzumab, cetuximab, depatuxizumab, zalutumumab, nexituzumab, panitumumab, emtricitab, nipocalimab, loliximab, broxatumomab, farucumab, socin-mituximab, datuximab, nasituximab, otilizumab monoclonal antibody, magetuximab, pertuzumab, trastuzumab, trastuzumab-drutilecan, trastuzumab-emtansine, trastuzumab-dulcamazine, pertuzumab, anifulurumab, epavacumab, ligoguzumab, omalizumab, daloizumab, fentolimumab, tetumumab, gevolizumab, canakinumab, daclizumab, basiliximab, dupilumab, mepolizumab, risorumab, benralizumab, clakizumab, ologizumab, selukuumab, siltuximab, sareluzumab, satarazumab, tocilizumab, REGN88, secukinumab, ustekinumab, brianuzumab, levofloxacin Birelizumab, troloxetine, ixekizumab, bimetuzumab, brodalumab, brazulizumab, guselkumab, risankizumab, tetrakizumab, migizumab, nemozumab, pesolimumab, relalizumab, nasopimumab, fasilumab, tanezumab, alirocumab, evolocumab, borcovacuzumab, pembrolizumab, bastimumab, carrelizumab, cemiplizumab, dotalimumab, palolizumab, cedizumab, spartalizumab, tislelizumab, pembrolizumab, nivolumab, atezolizumab, avelumab, envolimumab, durvalumab, bintrafuspα, denosumab, elotizumab, concilizumab, mastatumomab, infliximab, adalimumab, golimumab, certolizumab pegol, orizumab, brolucizumab, ranibizumab, bevacizumab, faretuzumab, ramucirumab, and caprolactam.

11. The pharmaceutical composition according to claim 4, wherein the soluble receptor or the soluble receptor contained in the Fc fusion protein of the soluble receptor is selected from the group consisting of TNF-α soluble receptor, VEGF soluble receptor, CTLA-4, interleukin 1 soluble receptor and LFA3 soluble receptor.

12. The pharmaceutical composition according to claim 4, wherein the Fc fusion protein of the soluble receptor is selected from aflibercept, abatacept, belatacept, rilonacept and alecept.

13. The pharmaceutical composition according to claim 1, wherein the drug is an anticancer drug.

14. The pharmaceutical composition according to claim 1, further comprising one or more selected from a buffer, a stabilizer and a surfactant.

15. The pharmaceutical composition of claim 14, wherein the buffer comprises one or more selected from the group consisting of malate, formate, citrate, acetate, propionate, pyridine, piperazine, cacodylate, succinate, 2-(N-morpholino)ethanesulfonic acid (MES), histidine, Tris, bis-Tris, phosphate, ethanolamine, carbonate, piperazine-N,N'-bis(2-ethanesulfonic acid) (PIPES), imidazole, BIS-TRIS propane, N,N-bis(2-hydroxyethyl)-2-aminoethanesulfonic acid (BES), 3-(N-morpholino)propanesulfonic acid (MOPS), hydroxyethylpiperazineethanesulfonic acid (HEPES), pyrophosphate, and triethanolamine; The stabilizer comprises one or more selected from the group consisting of carbohydrates, sugars or hydrates thereof, sugar alcohols or hydrates thereof, and amino acids; and The surfactant includes one or more nonionic surfactants selected from the group consisting of polyoxyethylene-sorbitan fatty acid esters, polyethylene-polypropylene glycol, polyoxyethylene-stearate, polyoxyethylene alkyl ether, polyoxyethylene-polyoxypropylene copolymer and sodium dodecyl sulfate (SDS).

16. The pharmaceutical composition according to claim 15, wherein the carbohydrate, the sugar or the sugar alcohol comprises one or more selected from the group consisting of trehalose or a hydrate thereof, sucrose, saccharin, glycerol, erythritol, threitol, xylitol, arabitol, ribitol, mannitol, sorbitol, galactitol, fucitol, iditol, inositol, heptyl alcohol, isomaltulose, maltitol, polyglucitol, cyclodextrin, hydroxypropyl cyclodextrin and glucose, and The amino acids include one or more selected from the group consisting of glutamine, glutamic acid, glycine, lysine, lysine dimer, leucine, methionine, valine, serine, selenomethionine, citrulline, arginine, asparagine, aspartic acid, ornithine, isoleucine, taurine, theanine, threonine, tryptophan, tyrosine, phenylalanine, proline, pyrrolysine, histidine and alanine.

17. The pharmaceutical composition of claim 1, wherein the pharmaceutical composition comprises a histidine buffer providing a pH of 5.5 ± 2.0, trehalose, and methionine.

18. The pharmaceutical composition of claim 1, wherein the pharmaceutical composition comprises a histidine buffer providing a pH of 5.5 ± 2.0, trehalose, methionine, and polysorbate.

19. The pharmaceutical composition according to claim 18, wherein the pharmaceutical composition comprises a histidine buffer providing a pH of 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.

Images