C3 complement inhibition polypeptide, preparation method thereof and application of C3 complement inhibition polypeptide in medicine

Abstract

The invention relates to a C3 complement inhibition polypeptide, a preparation method thereof and application of the C3 complement inhibition polypeptide in medicine. Specifically, the invention relates to a polypeptide capable of inhibiting C3 complement, which is capable of penetrating through a cell membrane or a body tissue barrier, in particular an ocular barrier, to a site where administration is desired. The invention also relates to a preparation method of the C3 complement inhibition polypeptide and application of the C3 complement inhibition polypeptide in treatment of ophthalmic diseases, especially treatment of age-related macular degeneration (AMD) secondary geographic atrophy (GA) and the like, and belongs to the field of biological medicine.

Description

Technical Field

[0001] The present invention relates to an anti-C3 complement factor polypeptide having the function of penetrating cell membranes or body tissue barriers, which can penetrate cell membranes or body tissue barriers to reach the desired site of administration, wherein the body tissue barrier is particularly an eye barrier. The present invention also relates to a preparation method of the anti-C3 complement factor polypeptide and its use in treating ophthalmic diseases, particularly treating age-related macular degeneration (AMD) and secondary geographic atrophy (GA), and belongs to the field of biomedicine. Background Art

[0002] Age-related macular degeneration (AMD) is an acquired retinal degeneration that causes severe central vision loss through the formation of non-neovascular (drusen and retinal pigment epithelial abnormalities) and neovascular disorders (choroidal neovascular membrane formation). AMD seriously affects the quality of life of patients, especially the elderly. It is an irreversible disease that leads to blindness. It is the third most common blinding eye disease in the world and the most common cause of irreversible blindness in the elderly.

[0003] Age-related macular degeneration (AMD) can be divided into wet and dry forms. Dry AMD accounts for about 80-90% of all AMD, but dry AMD may turn into wet AMD. Wet AMD, also known as vascular exudative age-related macular degeneration (AMD), is characterized by abnormal growth of blood vessels and infiltration of fluid and blood into the macula responsible for central vision. There is abnormal growth of new blood vessels behind the retina in the macular area. These new blood vessels are very fragile and prone to bleeding and fluid leakage, leading to edema and swelling of the macula, forming damage to the macula. As AMD progresses, the loss of retinal cells and underlying blood vessels in the macula leads to significant thinning and or atrophy of retinal tissue. Geographic atrophy associated with AMD leads to further irreversible vision loss in these patients.

[0004] The cause of wet AMD is complex, but choroidal neovascularization is its main feature. Currently, vascular endothelial growth factor (VEGF) inhibitors (such as ranibizumab, aflibercept, conbercept, etc.) are the leading drugs for the treatment of wet AMD, accounting for more than 95% of the global market share.

[0005] Current treatment ideas include delaying aging, protecting the optic nerve, cell therapy, anti-inflammation and inhibiting the complement system, among which complement inhibitors are the backbone of the current treatment of dry AMD; as of June 2023, only SYFOVRE (pegcetacoplan, C3 complement inhibitor) has been approved for marketing for the treatment of geographic atrophy (GA) secondary to age-related macular degeneration (AMD).

[0006] The complement system plays an important role in the pathogenesis of age-related macular degeneration (AMD). Studies have shown that complement system dysfunction may lead to the formation of drusen and GA, which are hallmarks of dry AMD. Proteins involved in the inflammatory and immune responses to drusen in the retina of patients with AMD include various components of the complement system (C3, C5, and C5b-9 complex). Multiple studies have shown a strong correlation between complement system gene variants and AMD, including complement factor H (CFH), C2, complement factor B (CFB), and C3 genes, and AMD patients also show higher levels of complement activity products, including C3d, Ba, C3a, C5a, SC5b-9, substrate proteins C3, C4, CFB, and regulatory factors CFH and complement factor D (CFD). C3 is the central molecule of the complement system. Three protein activation pathways, namely the classical pathway (CP), the lectin pathway (LP) and the alternative pathway (AP), converge on the complement system, leading to the activation of the terminal pathway (TP) of cell lysis, thereby triggering inflammatory or cytolytic immune responses. Abnormal complement activation causes the occurrence and development of AMD and even leads to geographic atrophy (GA).

[0007] Cell-penetrating peptides (CPPs) are a class of 3-35 amino acid polypeptides with typical characteristics of crossing cell membranes. They can effectively transport small molecule drugs, proteins, peptides, nucleic acid fragments, and nanocarriers (such as liposomes, polymer micelles, inorganic nanoparticles, etc.) across the cell membrane. CPPs have low toxicity and no immunogenicity, and can connect / carry a wide range of cargoes. Summary of the invention

[0008] The present invention develops functional polypeptides that have the ability to penetrate the membrane and serve as C3 complement inhibitors. These functional peptides can be introduced into the eye in a non-invasive manner, thereby achieving the effect of continuously, stably and effectively treating diseases such as age-related macular degeneration (AMD), diabetic macular edema (DME), and macular edema caused by retinal vein occlusion, and thus has good application value.

[0009] Therefore, the purpose of the present invention is to provide a peptide having the function of penetrating cell membranes or body tissue barriers and its application, so as to provide an effective treatment solution for diseases such as age-related macular degeneration (AMD) and diabetic macular edema (DME) caused by retinal microvascular disorders.

[0010] In one aspect, the present invention provides a polypeptide comprising or consisting of the following amino acid sequence:

[0011] X 1 ICVX 2 QDJ 3AHRCI (SEQ ID NO: 1), wherein

[0012] X 1 represents D-tyrosine;

[0013] X 2 is tryptophan or a tryptophan analogue selected from the group consisting of:

[0014] a) N-methyltryptophan;

[0015] b) tryptophan having a C1-C4 alkyl substituent at the 5-position of the indole ring;

[0016] c) 5-fluorotryptophan; or

[0017] d) 6-fluorotryptophan;

[0018] X 3 represents sarcosine;

[0019] A disulfide bond is formed between the cysteine ​​residues at positions 3 and 13.

[0020] In another aspect, the present invention provides a polypeptide comprising, or consisting of, the following amino acid sequence:

[0021] X 4 CVX 2 QDWGAHRCT (SEQ ID NO: 11), wherein

[0022] X 2 is tryptophan or a tryptophan analogue selected from

[0023] a) N-methyltryptophan;

[0024] b) tryptophan having a C1-C4 alkyl substituent at the 5-position of the indole ring;

[0025] c) 5-fluorotryptophan; or

[0026] d) 6-fluorotryptophan;

[0027] X 4 indicates acetylated isoleucine;

[0028] A disulfide bond is formed between the 2nd and 12th cysteines.

[0029] In another aspect, the present invention provides a fusion polypeptide comprising:

[0030] A) a polypeptide represented by SEQ ID NO: 1,

[0031] X 1 ICVX2 QDJ 3 AHRCI (SEQ ID NO: 1), wherein

[0032] X 1 represents D-tyrosine;

[0033] X 2 Selected from tryptophan or a tryptophan analog, wherein the tryptophan analog is selected from:

[0034] a) N-methyltryptophan;

[0035] b) tryptophan having a C1-C4 alkyl substituent at the 5-position of the indole ring;

[0036] c) 5-fluorotryptophan; or

[0037] d) 6-fluorotryptophan;

[0038] X 3 represents sarcosine;

[0039] wherein a disulfide bond is formed between the cysteine ​​residues at positions 3 and 13; and

[0040] B) A polypeptide comprising any of the following amino acid sequences:

[0041] GRKKRRQRRRPQ (SEQ ID NO: 21);

[0042] RQIKIWFQNRRMKWKK (SEQ ID NO: 22);

[0043] GRKKRRQRRRPQ (SEQ ID NO: 23);

[0044] YGRKKRRQRRR (SEQ ID NO: 24);

[0045] GYGRKKRRQRRR (SEQ ID NO: 25);

[0046] GGYGRKKRRQRRR (SEQ ID NO: 26); or

[0047] GGGYGRKKRRQRRR (SEQ ID NO: 27);

[0048] Optionally, the polypeptide shown in any one of SEQ ID NO: 21 to SEQ ID NO: 27 has or does not have C-terminal amidation;

[0049] Preferably, the polypeptide shown in any one of SEQ ID NO: 21 to SEQ ID NO: 27 has a membrane-penetrating function;

[0050] Preferably, the polypeptide shown in any one of SEQ ID NO:21 to SEQ ID NO:27 is located at the C-terminus of the polypeptide shown in SEQ ID NO:1.

[0051] In an embodiment of the fusion polypeptide of the present invention, the polypeptide shown in SEQ ID NO: 1 is directly linked to any polypeptide having transmembrane properties shown in SEQ ID NO: 21 to SEQ ID NO: 27, or is linked via a linker.

[0052] In an embodiment of the fusion polypeptide of the present invention, the polypeptide shown in SEQ ID NO: 1 is connected to any polypeptide with transmembrane properties shown in SEQ ID NO: 21 to SEQ ID NO: 27 through a linker, and optionally, the linker is a fragment shown in AEEA.

[0053] AEEA represents 8-amino-3,6-dioxaoctanoyl (-NH-CH 2 CH 2 OCH 2 CH 2 -C(=O)-).

[0054] In another aspect, the present invention provides a fusion polypeptide comprising:

[0055] A) a polypeptide represented by SEQ ID NO: 11,

[0056] X 4 CVX 2 QDWGAHRCT (SEQ ID NO: 11), wherein

[0057] X 2 is tryptophan or a tryptophan analogue selected from

[0058] a) N-methyltryptophan;

[0059] b) tryptophan having a C1-C4 alkyl substituent at the 5-position of the indole ring;

[0060] c) 5-fluorotryptophan; or

[0061] d) 6-fluorotryptophan;

[0062] X 4 indicates acetylated isoleucine;

[0063] wherein a disulfide bond is formed between the cysteine ​​residues at positions 2 and 12; and

[0064] B) a polypeptide comprising the following amino acid sequence:

[0065] GRKKRRQRRRPQ (SEQ ID NO: 21);

[0066] RQIKIWFQNRRMKWKK (SEQ ID NO: 22);

[0067] GRKKRRQRRRPQ (SEQ ID NO: 23);

[0068] YGRKKRRQRRR (SEQ ID NO: 24);

[0069] GYGRKKRRQRRR (SEQ ID NO: 25);

[0070] GGYGRKKRRQRRR (SEQ ID NO: 26); or

[0071] GGGYGRKKRRQRRR (SEQ ID NO: 27);

[0072] Optionally, the polypeptide shown in any one of SEQ ID NO: 21 to SEQ ID NO: 27 has or does not have C-terminal amidation;

[0073] Preferably, the polypeptide shown in any one of SEQ ID NO: 21 to SEQ ID NO: 27 has a membrane-penetrating function;

[0074] Preferably, the polypeptide shown in any one of SEQ ID NO:21 to SEQ ID NO:27 is located at the C-terminus of the polypeptide shown in SEQ ID NO:11.

[0075] In an embodiment of the fusion polypeptide of the present invention, the polypeptide represented by SEQ ID NO: 11 is directly linked to any polypeptide having transmembrane properties represented by SEQ ID NO: 21 to SEQ ID NO: 27, or is linked via a linker.

[0076] In an embodiment of the fusion polypeptide of the present invention, the polypeptide shown in SEQ ID NO: 11 is connected to any polypeptide with transmembrane properties shown in SEQ ID NO: I to SEQ ID NO: VII through a linker, and optionally, the linker is a fragment shown in AEEA.

[0077] AEEA represents 8-amino-3,6-dioxaoctanoyl (-NH-CH 2 CH 2 OCH 2 CH 2 -C(=O)-).

[0078] In an embodiment of the polypeptide or fusion polypeptide of the present invention, the polypeptide or fusion polypeptide has an acetylation modification at the N-terminus and / or an amidation modification at the C-terminus.

[0079] The polypeptide or fusion polypeptide provided by the present invention can function as a C3 complement inhibitor and can penetrate cell membranes or body tissue barriers.

[0080] Thus, in another aspect, the present invention provides a pharmaceutical composition comprising a polypeptide or fusion polypeptide as described herein, and one or more pharmaceutically acceptable carriers or excipients.

[0081] In another aspect, the present invention provides a nucleic acid molecule encoding a polypeptide or fusion polypeptide of the present invention.

[0082] In another aspect, the present invention provides a vector comprising a nucleic acid molecule of the present invention.

[0083] In another aspect, the present invention provides a host cell comprising a nucleic acid molecule or a vector of the present invention.

[0084] In another aspect, the present invention provides the polypeptide or fusion polypeptide of the present invention or a pharmaceutical composition comprising the same, for use in treating diseases, especially eye diseases.

[0085] In another aspect, the present invention provides use of the polypeptide or fusion polypeptide of the present invention or a pharmaceutical composition comprising the same in the preparation of a medicament for treating a disease, especially an eye disease.

[0086] In a specific embodiment of the invention, the ocular disease is selected from age-related macular degeneration (AMD), diabetic macular edema (DME), inflammatory eye diseases, optic neuropathy eye diseases and ocular microangiopathy-related disorders.

[0087] In some embodiments of the present invention, the ocular microangiopathy-related disorder is selected from diabetic retinopathy (DR), diabetic macular edema (DME), retinal vein occlusion (RVO), central retinal vein occlusion (CRVO), macular degeneration, retinopathy of prematurity (ROP), neovascular glaucoma, retinitis pigmentosa (RP), proliferative and non-proliferative retinopathy, retinal angiomatous proliferation, macular telangiectasia, ischemic retinopathy, iris neovascularization, intraocular neovascularization, corneal neovascularization, retinal neovascularization, polypoidal choroidal vasculopathy (PCV), choroidal neovascularization, retinal degeneration, diabetic complications of retinal vascular permeability associated with diabetic retinopathy and diabetic macular edema; in particular, selected from diabetic retinopathy (DR), diabetic macular edema (DME), retinal vein occlusion (RVO), central retinal vein occlusion (CRVO) and wet age-related macular degeneration (wet AMD).

[0088] In another aspect, the present invention provides a method for treating a disease, especially an ocular disease, comprising administering to a subject in need thereof a therapeutically effective amount of a polypeptide or fusion polypeptide according to the present invention or a pharmaceutical composition comprising the same. DETAILED DESCRIPTION

[0089] Specifically, the present invention provides a polypeptide having the amino acid sequence shown in SEQ ID NO: 1; or having at least 95%, preferably at least 94%, at least 93%, at least 92%, at least 91%, at least 90%, at least 89%, at least 87%, at least 86%, at least 85%, at least 84%, at least 83%, at least 82%, at least 81%, at least 80%, at least 79%, at least 78%, at least 77%, at least 76%, at least 75% sequence identity with the amino acid sequence shown in SEQ ID NO: 1; or having at most 3, preferably at most 2 or at most 1 amino acid sequence substitutions, deletions or additions compared to SEQ ID NO: 1:

[0090] X 1 ICVX 2 QDJ 3 AHRCI (SEQ ID NO: 1), wherein

[0091] X 1 represents D-tyrosine;

[0092] X 2 is tryptophan or a tryptophan analogue selected from the group consisting of:

[0093] a) N-methyltryptophan;

[0094] b) tryptophan having a C1-C4 alkyl substituent at the 5-position of the indole ring;

[0095] c) 5-fluorotryptophan; or

[0096] d) 6-fluorotryptophan;

[0097] X 3 represents sarcosine;

[0098] A disulfide bond is formed between the cysteine ​​residues at positions 3 and 13.

[0099] In a specific embodiment, the present invention provides a polypeptide having the amino acid sequence shown in SEQ ID NO: 2; or having at least 95%, preferably at least 94%, at least 93%, at least 92%, at least 91%, at least 90%, at least 89%, at least 87%, at least 86%, at least 85%, at least 84%, at least 83%, at least 82%, at least 81%, at least 80%, at least 79%, at least 78%, at least 77%, at least 76%, at least 75% sequence identity with the amino acid sequence shown in SEQ ID NO: 2; or having at most 3, preferably at most 2 or at most 1 amino acid sequence substitutions, deletions or additions compared to SEQ ID NO: 2:

[0100] X 1 ICVX 2 QDJ 3 AHRCIGRKKRRQRRRPQ(SEQ ID NO:2),

[0101] in

[0102] X 1 represents D-tyrosine;

[0103] X 2 is tryptophan or a tryptophan analogue selected from the group consisting of:

[0104] a) N-methyltryptophan;

[0105] b) tryptophan having a C1-C4 alkyl substituent at the 5-position of the indole ring;

[0106] c) 5-fluorotryptophan; or

[0107] d) 6-fluorotryptophan;

[0108] X 3 represents sarcosine;

[0109] A disulfide bond is formed between the 3rd and 13th cysteines;

[0110] wherein the C-terminus of SEQ ID NO: 2 is amidated.

[0111] In a specific embodiment, the present invention provides a polypeptide having the amino acid sequence shown in SEQ ID NO:3 as follows; or having at least 95%, preferably at least 94%, at least 93%, at least 92%, at least 91%, at least 90%, at least 89%, at least 87%, at least 86%, at least 85%, at least 84%, at least 83%, at least 82%, at least 81%, at least 80%, at least 79%, at least 78%, at least 77%, at least 76%, at least 75% sequence identity with the amino acid sequence shown in SEQ ID NO:3; or having at most 3, preferably at most 2 or at most 1 amino acid sequence substitutions, deletions or additions compared to SEQ ID NO:3:

[0112] X 1 ICVX 2 QDJ 3 AHRCIX 5 GRKKRRQRRRPQ (SEQ ID NO: 3), wherein

[0113] X 1 represents D-tyrosine;

[0114] X 2 is tryptophan or a tryptophan analogue selected from the group consisting of:

[0115] a) N-methyltryptophan;

[0116] b) tryptophan having a C1-C4 alkyl substituent at the 5-position of the indole ring;

[0117] c) 5-fluorotryptophan; or

[0118] d) 6-fluorotryptophan;

[0119] X 3 represents sarcosine;

[0120] X 5 AEEA, 8-amino-3,6-dioxaoctanoyl (-NH-CH 2 CH 2 OCH 2 CH 2 -C(=O)-);

[0121] A disulfide bond is formed between the 3rd and 13th cysteines;

[0122] wherein the C-terminus of SEQ ID NO: 3 is amidated.

[0123] In a specific embodiment, the present invention provides a polypeptide having the amino acid sequence shown in SEQ ID NO:4 as follows; or having at least 95%, preferably at least 94%, at least 93%, at least 92%, at least 91%, at least 90%, at least 89%, at least 87%, at least 86%, at least 85%, at least 84%, at least 83%, at least 82%, at least 81%, at least 80%, at least 79%, at least 78%, at least 77%, at least 76%, at least 75% sequence identity with the amino acid sequence shown in SEQ ID NO:4; or having at most 3, preferably at most 2 or at most 1 amino acid sequence substitutions, deletions or additions compared to SEQ ID NO:4:

[0124] X 1 ICVX 2 QDJ 3 AHRCI RQIKIWFQNRRMKWKK (SEQ ID NO: 4), wherein

[0125] X 1 represents D-tyrosine;

[0126] X 2 is tryptophan or a tryptophan analogue selected from the group consisting of:

[0127] a) N-methyltryptophan;

[0128] b) tryptophan having a C1-C4 alkyl substituent at the 5-position of the indole ring;

[0129] c) 5-fluorotryptophan; or

[0130] d) 6-fluorotryptophan;

[0131] X 3 represents sarcosine;

[0132] A disulfide bond is formed between the 3rd and 13th cysteines;

[0133] wherein the C-terminus of SEQ ID NO: 4 is amidated.

[0134] In a specific embodiment, the present invention provides a polypeptide having the amino acid sequence shown in SEQ ID NO:5; or having at least 95%, preferably at least 94%, at least 93%, at least 92%, at least 91%, at least 90%, at least 89%, at least 87%, at least 86%, at least 85%, at least 84%, at least 83%, at least 82%, at least 81%, at least 80%, at least 79%, at least 78%, at least 77%, at least 76%, at least 75% sequence identity with the amino acid sequence shown in SEQ ID NO:5; or having at most 3, preferably at most 2 or at most 1 amino acid sequence substitutions, deletions or additions compared to SEQ ID NO:5:

[0135] X 1 ICVX 2 QDJ 3 AHRCI X 5 RQIKIWFQNRRMKWKK(SEQ ID NO:5),

[0136] in

[0137] X 1 represents D-tyrosine;

[0138] X 2 is tryptophan or a tryptophan analogue selected from the group consisting of:

[0139] a) N-methyltryptophan;

[0140] b) tryptophan having a C1-C4 alkyl substituent at the 5-position of the indole ring;

[0141] c) 5-fluorotryptophan; or

[0142] d) 6-fluorotryptophan;

[0143] X 3 represents sarcosine;

[0144] X 5 AEEA; 8-amino-3,6-dioxaoctanoyl (-NH-CH 2 CH 2 OCH 2 CH 2 -C(=O)-);

[0145] A disulfide bond is formed between the 3rd and 13th cysteines;

[0146] wherein the C-terminus of SEQ ID NO: 5 is amidated.

[0147] In a specific embodiment, the present invention provides a polypeptide having the amino acid sequence shown in SEQ ID NO:6 as follows; or having at least 95%, preferably at least 94%, at least 93%, at least 92%, at least 91%, at least 90%, at least 89%, at least 87%, at least 86%, at least 85%, at least 84%, at least 83%, at least 82%, at least 81%, at least 80%, at least 79%, at least 78%, at least 77%, at least 76%, at least 75% sequence identity with the amino acid sequence shown in SEQ ID NO:6; or having at most 3, preferably at most 2 or at most 1 amino acid sequence substitutions, deletions or additions compared to SEQ ID NO:6:

[0148] X 1 ICVX 2 QDJ 3 AHRCIX 5 GRKKRRQRRRPQ (SEQ ID NO: 6), wherein

[0149] X 1 represents D-tyrosine;

[0150] X 2 is tryptophan or a tryptophan analogue selected from the group consisting of:

[0151] a) N-methyltryptophan;

[0152] b) tryptophan having a C1-C4 alkyl substituent at the 5-position of the indole ring;

[0153] c) 5-fluorotryptophan; or

[0154] d) 6-fluorotryptophan;

[0155] X 3 represents sarcosine;

[0156] X 5 AEEA, 8-amino-3,6-dioxaoctanoyl (-NH-CH 2 CH 2 OCH 2 CH 2 -C(=O)-);

[0157] A disulfide bond is formed between the cysteine ​​residues at positions 3 and 13.

[0158] In a specific embodiment, the present invention provides a polypeptide having the amino acid sequence shown in SEQ ID NO:7 as follows; or having at least 95%, preferably at least 94%, at least 93%, at least 92%, at least 91%, at least 90%, at least 89%, at least 87%, at least 86%, at least 85%, at least 84%, at least 83%, at least 82%, at least 81%, at least 80%, at least 79%, at least 78%, at least 77%, at least 76%, at least 75% sequence identity with the amino acid sequence shown in SEQ ID NO:7; or having up to 3, preferably up to 2 or up to 1 amino acid sequence substitutions, deletions or additions compared to SEQ ID NO:7:

[0159] X 1 ICVX 2 QDJ 3 AHRCIX 5 YGRKKRRQRRR (SEQ ID NO: 7), wherein

[0160] X 1 represents D-tyrosine;

[0161] X 2 is tryptophan or a tryptophan analogue selected from the group consisting of:

[0162] a) N-methyltryptophan;

[0163] b) tryptophan having a C1-C4 alkyl substituent at the 5-position of the indole ring;

[0164] c) 5-fluorotryptophan; or

[0165] d) 6-fluorotryptophan;

[0166] X 3 represents sarcosine;

[0167] X5 represents AEEA, that is, 8-amino-3,6-dioxaoctanoyl (-NH-CH 2 CH 2 OCH 2 CH 2 -C(=O)-);

[0168] A disulfide bond is formed between the cysteine ​​residues at positions 3 and 13.

[0169] In a specific embodiment, the present invention provides a polypeptide having the amino acid sequence shown in SEQ ID NO: 8; or having at least 95%, preferably at least 94%, at least 93%, at least 92%, at least 91%, at least 90%, at least 89%, at least 87%, at least 86%, at least 85%, at least 84%, at least 83%, at least 82%, at least 81%, at least 80%, at least 79%, at least 78%, at least 77%, at least 76%, at least 75% sequence identity with the amino acid sequence shown in SEQ ID NO: 8; or having at most 3, preferably at most 2 or at most 1 amino acid sequence substitutions, deletions or additions compared to SEQ ID NO: 8:

[0170] X 1 ICVX 2 QDJ 3 AHRCIX5GYGRKKRRQRRR (SEQ ID NO: 8), wherein

[0171] X 1 represents D-tyrosine;

[0172] X 2 is tryptophan or a tryptophan analogue selected from the group consisting of:

[0173] a) N-methyltryptophan;

[0174] b) tryptophan having a C1-C4 alkyl substituent at the 5-position of the indole ring;

[0175] c) 5-fluorotryptophan; or

[0176] d) 6-fluorotryptophan;

[0177] X 3 represents sarcosine;

[0178] X 5 AEEA; 8-amino-3,6-dioxaoctanoyl (-NH-CH 2 CH 2 OCH 2 CH 2 -C(=O)-);

[0179] A disulfide bond is formed between the 3rd and 13th cysteines;

[0180] wherein the C-terminus of SEQ ID NO: 8 is amidated.

[0181] In a specific embodiment, the present invention provides a polypeptide having the amino acid sequence shown in SEQ ID NO:9 as follows; or having at least 95%, preferably at least 94%, at least 93%, at least 92%, at least 91%, at least 90%, at least 89%, at least 87%, at least 86%, at least 85%, at least 84%, at least 83%, at least 82%, at least 81%, at least 80%, at least 79%, at least 78%, at least 77%, at least 76%, at least 75% sequence identity with the amino acid sequence shown in SEQ ID NO:9; or having at most 3, preferably at most 2 or at most 1 amino acid sequence substitutions, deletions or additions compared to SEQ ID NO:9:

[0182] X 1 ICVX 2 QDJ 3 AHRCIX 5 GGYGRKKRRQRRR (SEQ ID NO: 9), wherein

[0183] X 1 represents D-tyrosine;

[0184] X 2 is tryptophan or a tryptophan analogue selected from the group consisting of:

[0185] a) N-methyltryptophan;

[0186] b) tryptophan having a C1-C4 alkyl substituent at the 5-position of the indole ring;

[0187] c) 5-fluorotryptophan; or

[0188] d) 6-fluorotryptophan;

[0189] X 3 represents sarcosine;

[0190] X5 represents AEEA, i.e. 8-amino-3,6-dioxaoctanoyl (-NH-CH 2 CH 2 OCH 2 CH 2 -C(=O)-);

[0191] A disulfide bond is formed between the 3rd and 13th cysteines;

[0192] wherein the C-terminus of SEQ ID NO: 9 is amidated.

[0193] In a specific embodiment, the present invention provides a polypeptide having the amino acid sequence shown in SEQ ID NO: 10; or having at least 95%, preferably at least 94%, at least 93%, at least 92%, at least 91%, at least 90%, at least 89%, at least 87%, at least 86%, at least 85%, at least 84%, at least 83%, at least 82%, at least 81%, at least 80%, at least 79%, at least 78%, at least 77%, at least 76%, at least 75% sequence identity with the amino acid sequence shown in SEQ ID NO: 10; or having at most 3, preferably at most 2 or at most 1 amino acid sequence substitutions, deletions or additions compared to SEQ ID NO: 10:

[0194] X 1 ICVX 2 QDJ 3 AHRCIX 5 GGGYGRKKRRQRRR (SEQ ID NO: 10), wherein

[0195] X 1 represents D-tyrosine;

[0196] X 2 is tryptophan or a tryptophan analogue selected from the group consisting of:

[0197] a) N-methyltryptophan;

[0198] b) tryptophan having a C1-C4 alkyl substituent at the 5-position of the indole ring;

[0199] c) 5-fluorotryptophan; or

[0200] d) 6-fluorotryptophan;

[0201] X 3 represents sarcosine;

[0202] X5 represents AEEA, i.e. 8-amino-3,6-dioxaoctanoyl (-NH-CH 2 CH 2 OCH 2 CH 2 -C(=O)-);

[0203] A disulfide bond is formed between the 3rd and 13th cysteines;

[0204] wherein the C-terminus of SEQ ID NO: 10 is amidated.

[0205] In a specific embodiment, the present invention provides a polypeptide having the amino acid sequence shown in SEQ ID NO: 11 as follows; or having at least 95%, preferably at least 94%, at least 93%, at least 92%, at least 91%, at least 90%, at least 89%, at least 87%, at least 86%, at least 85%, at least 84%, at least 83%, at least 82%, at least 81%, at least 80%, at least 79%, at least 78%, at least 77%, at least 76%, at least 75% sequence identity with the amino acid sequence shown in SEQ ID NO: 11; or having at most 3, preferably at most 2 or at most 1 amino acid sequence substitutions, deletions or additions compared to SEQ ID NO: 11:

[0206] X 4 CVX 2 QDWGAHRCT (SEQ ID NO: 11),

[0207] in

[0208] X 2 is tryptophan or a tryptophan analogue selected from

[0209] a) N-methyltryptophan;

[0210] b) tryptophan having a C1-C4 alkyl substituent at the 5-position of the indole ring;

[0211] c) 5-fluorotryptophan; or

[0212] d) 6-fluorotryptophan;

[0213] X 4 indicates acetylated isoleucine;

[0214] A disulfide bond is formed between the 2nd and 12th cysteines.

[0215] In a specific embodiment, the present invention provides a polypeptide having the amino acid sequence shown in SEQ ID NO: 12 as follows; or having at least 95%, preferably at least 94%, at least 93%, at least 92%, at least 91%, at least 90%, at least 89%, at least 87%, at least 86%, at least 85%, at least 84%, at least 83%, at least 82%, at least 81%, at least 80%, at least 79%, at least 78%, at least 77%, at least 76%, at least 75% sequence identity with the amino acid sequence shown in SEQ ID NO: 12; or having at most 3, preferably at most 2 or at most 1 amino acid sequence substitutions, deletions or additions compared to SEQ ID NO: 12:

[0216] X 4CVX 2 QDWGAHRCTGRKKRRQRRRPQ (SEQ ID NO: 12), wherein

[0217] X 2 is tryptophan or a tryptophan analogue selected from

[0218] a) N-methyltryptophan;

[0219] b) tryptophan having a C1-C4 alkyl substituent at the 5-position of the indole ring;

[0220] c) 5-fluorotryptophan; or

[0221] d) 6-fluorotryptophan;

[0222] X 4 indicates acetylated isoleucine;

[0223] A disulfide bond is formed between the 2nd and 12th cysteines;

[0224] wherein the C-terminus of SEQ ID NO: 12 is amidated.

[0225] In a specific embodiment, the present invention provides a polypeptide having the amino acid sequence shown in SEQ ID NO: 13 as follows; or having at least 95%, preferably at least 94%, at least 93%, at least 92%, at least 91%, at least 90%, at least 89%, at least 87%, at least 86%, at least 85%, at least 84%, at least 83%, at least 82%, at least 81%, at least 80%, at least 79%, at least 78%, at least 77%, at least 76%, at least 75% sequence identity with the amino acid sequence shown in SEQ ID NO: 13; or having at most 3, preferably at most 2 or at most 1 amino acid sequence substitutions, deletions or additions compared to SEQ ID NO: 13:

[0226] X 4 CVX 2 QDWGAHRCTX5GRKKRRQRRRPQ (SEQ ID NO: 13), wherein

[0227] X 2 is tryptophan or a tryptophan analogue selected from

[0228] a) N-methyltryptophan;

[0229] b) tryptophan having a C1-C4 alkyl substituent at the 5-position of the indole ring;

[0230] c) 5-fluorotryptophan; or

[0231] d) 6-fluorotryptophan;

[0232] X 4 indicates acetylated isoleucine;

[0233] X 5 AEEA, 8-amino-3,6-dioxaoctanoyl (-NH-CH 2 CH 2 OCH 2 CH 2 -C(=O)-);

[0234] A disulfide bond is formed between the 2nd and 12th cysteines;

[0235] wherein the C-terminus of SEQ ID NO: 13 is amidated.

[0236] In a specific embodiment, the present invention provides a polypeptide having the amino acid sequence shown in SEQ ID NO: 14 as follows; or having at least 95%, preferably at least 94%, at least 93%, at least 92%, at least 91%, at least 90%, at least 89%, at least 87%, at least 86%, at least 85%, at least 84%, at least 83%, at least 82%, at least 81%, at least 80%, at least 79%, at least 78%, at least 77%, at least 76%, at least 75% sequence identity with the amino acid sequence shown in SEQ ID NO: 14; or having at most 3, preferably at most 2 or at most 1 amino acid sequence substitutions, deletions or additions compared to SEQ ID NO: 14:

[0237] X 4 CVX 2 QDWGAHRCTRQIKIWFQNRRMKWKK (SEQ ID NO: 14), wherein

[0238] X 2 is tryptophan or a tryptophan analogue selected from

[0239] a) N-methyltryptophan;

[0240] b) tryptophan having a C1-C4 alkyl substituent at the 5-position of the indole ring;

[0241] c) 5-fluorotryptophan; or

[0242] d) 6-fluorotryptophan;

[0243] X 4 indicates acetylated isoleucine;

[0244] A disulfide bond is formed between the 2nd and 12th cysteines;

[0245] wherein the C-terminus of SEQ ID NO: 14 is amidated.

[0246] In a specific embodiment, the present invention provides a polypeptide having the amino acid sequence shown in SEQ ID NO: 15 as follows; or having at least 95%, preferably at least 94%, at least 93%, at least 92%, at least 91%, at least 90%, at least 89%, at least 87%, at least 86%, at least 85%, at least 84%, at least 83%, at least 82%, at least 81%, at least 80%, at least 79%, at least 78%, at least 77%, at least 76%, at least 75% sequence identity with the amino acid sequence shown in SEQ ID NO: 15; or having up to 3, preferably up to 2 or up to 1 amino acid sequence substitutions, deletions or additions compared to SEQ ID NO: 15:

[0247] X 4 CVX 2 QDWGAHRCTX 5 RQIKIWFQNRRMKWKK(SEQ ID NO:15),

[0248] in

[0249] X 2 is tryptophan or a tryptophan analogue selected from

[0250] a) N-methyltryptophan;

[0251] b) tryptophan having a C1-C4 alkyl substituent at the 5-position of the indole ring;

[0252] c) 5-fluorotryptophan; or

[0253] d) 6-fluorotryptophan;

[0254] X 5 AEEA, 8-amino-3,6-dioxaoctanoyl (-NH-CH 2 CH 2 OCH 2 CH 2 -C(=O)-);

[0255] X 4 indicates acetylated isoleucine;

[0256] A disulfide bond is formed between the 2nd and 12th cysteines;

[0257] wherein the C-terminus of SEQ ID NO: 15 is amidated.

[0258] In a specific embodiment, the present invention provides a polypeptide having the amino acid sequence shown in SEQ ID NO: 16 as follows; or having at least 95%, preferably at least 94%, at least 93%, at least 92%, at least 91%, at least 90%, at least 89%, at least 87%, at least 86%, at least 85%, at least 84%, at least 83%, at least 82%, at least 81%, at least 80%, at least 79%, at least 78%, at least 77%, at least 76%, at least 75% sequence identity with the amino acid sequence shown in SEQ ID NO: 16; or having at most 3, preferably at most 2 or at most 1 amino acid sequence substitutions, deletions or additions compared to SEQ ID NO: 16:

[0259] X 4 CVX 2 QDWGAHRCTX 5 GRKKRRQRRRPQ (SEQ ID NO: 16), wherein

[0260] X 2 is tryptophan or a tryptophan analogue selected from

[0261] a) N-methyltryptophan;

[0262] b) tryptophan having a C1-C4 alkyl substituent at the 5-position of the indole ring;

[0263] c) 5-fluorotryptophan; or

[0264] d) 6-fluorotryptophan;

[0265] X 4 indicates acetylated isoleucine;

[0266] X 5 AEEA, 8-amino-3,6-dioxaoctanoyl (-NH-CH 2 CH 2 OCH 2 CH 2 -C(=O)-);

[0267] A disulfide bond is formed between the 2nd and 12th cysteines.

[0268] In a specific embodiment, the present invention provides a polypeptide having the amino acid sequence shown in SEQ ID NO: 17 as follows; or having at least 95%, preferably at least 94%, at least 93%, at least 92%, at least 91%, at least 90%, at least 89%, at least 87%, at least 86%, at least 85%, at least 84%, at least 83%, at least 82%, at least 81%, at least 80%, at least 79%, at least 78%, at least 77%, at least 76%, at least 75% sequence identity with the amino acid sequence shown in SEQ ID NO: 17; or having at most 3, preferably at most 2 or at most 1 amino acid sequence substitutions, deletions or additions compared to SEQ ID NO: 17:

[0269] X 4 CVX 2 QDWGAHRCTX 5 YGRKKRRQRRR (SEQ ID NO: 17), wherein

[0270] X 2 is tryptophan or a tryptophan analogue selected from

[0271] a) N-methyltryptophan;

[0272] b) tryptophan having a C1-C4 alkyl substituent at the 5-position of the indole ring;

[0273] c) 5-fluorotryptophan; or

[0274] d) 6-fluorotryptophan;

[0275] X 4 indicates acetylated isoleucine;

[0276] X 5 AEEA, 8-amino-3,6-dioxaoctanoyl (-NH-CH 2 CH 2 OCH 2 CH 2 -C(=O)-);

[0277] A disulfide bond is formed between the 2nd and 12th cysteines;

[0278] wherein the C-terminus of SEQ ID NO: 17 is amidated.

[0279] In a specific embodiment, the present invention provides a polypeptide having the amino acid sequence shown in SEQ ID NO: 18 as follows; or having at least 95%, preferably at least 94%, at least 93%, at least 92%, at least 91%, at least 90%, at least 89%, at least 87%, at least 86%, at least 85%, at least 84%, at least 83%, at least 82%, at least 81%, at least 80%, at least 79%, at least 78%, at least 77%, at least 76%, at least 75% sequence identity with the amino acid sequence shown in SEQ ID NO: 18; or having up to 3, preferably up to 2 or up to 1 amino acid sequence substitutions, deletions or additions compared to SEQ ID NO: 18:

[0280] X 4 CVX 2 QDWGAHRCTX 5 GYGRKKRRQRRR (SEQ ID NO: 18), wherein

[0281] X 2 is tryptophan or a tryptophan analogue selected from

[0282] a) N-methyltryptophan;

[0283] b) tryptophan having a C1-C4 alkyl substituent at the 5-position of the indole ring;

[0284] c) 5-fluorotryptophan; or

[0285] d) 6-fluorotryptophan;

[0286] X 4 indicates acetylated isoleucine;

[0287] X5 represents AEEA, i.e. 8-amino-3,6-dioxaoctanoyl (-NH-CH 2 CH 2 OCH 2 CH 2 -C(=O)-);

[0288] A disulfide bond is formed between the 2nd and 12th cysteines;

[0289] wherein the C-terminus of SEQ ID NO: 18 is amidated.

[0290] In a specific embodiment, the present invention provides a polypeptide having the amino acid sequence shown in SEQ ID NO: 19 as follows; or having at least 95%, preferably at least 94%, at least 93%, at least 92%, at least 91%, at least 90%, at least 89%, at least 87%, at least 86%, at least 85%, at least 84%, at least 83%, at least 82%, at least 81%, at least 80%, at least 79%, at least 78%, at least 77%, at least 76%, at least 75% sequence identity with the amino acid sequence shown in SEQ ID NO: 19; or having up to 3, preferably up to 2 or up to 1 amino acid sequence substitutions, deletions or additions compared to SEQ ID NO: 19:

[0291] X 4 CVX 2 QDWGAHRCTX 5 GGYGRKKRRQRRR (SEQ ID NO: 19), wherein

[0292] X 2 is tryptophan or a tryptophan analogue selected from

[0293] a) N-methyltryptophan;

[0294] b) tryptophan having a C1-C4 alkyl substituent at the 5-position of the indole ring;

[0295] c) 5-fluorotryptophan; or

[0296] d) 6-fluorotryptophan;

[0297] X 4 indicates acetylated isoleucine;

[0298] X 5 AEEA stands for 8-amino-3,6-dioxaoctanoyl (-NH-CH 2 CH 2 OCH 2 CH 2 -C(=O)-);

[0299] A disulfide bond is formed between the 2nd and 12th cysteines;

[0300] wherein the C-terminus of SEQ ID NO: 19 is amidated.

[0301] In a specific embodiment, the present invention provides a polypeptide having the amino acid sequence shown in SEQ ID NO: 20; or having at least 95%, preferably at least 94%, at least 93%, at least 92%, at least 91%, at least 90%, at least 89%, at least 87%, at least 86%, at least 85%, at least 84%, at least 83%, at least 82%, at least 81%, at least 80%, at least 79%, at least 78%, at least 77%, at least 76%, at least 75% sequence identity with the amino acid sequence shown in SEQ ID NO: 20; or having up to 3, preferably up to 2 or up to 1 amino acid sequence substitutions, deletions or additions compared to SEQ ID NO: 20:

[0302] X 4 CVX 2 QDWGAHRCTX 5 GGGYGRKKRRQRRR (SEQ ID NO: 20), wherein

[0303] X 2 is tryptophan or a tryptophan analogue selected from

[0304] a) N-methyltryptophan;

[0305] b) tryptophan having a C1-C4 alkyl substituent at the 5-position of the indole ring;

[0306] c) 5-fluorotryptophan; or

[0307] d) 6-fluorotryptophan;

[0308] X 4 indicates acetylated isoleucine;

[0309] X 5 AEEA stands for 8-amino-3,6-dioxaoctanoyl (-NH-CH 2 CH 2 OCH 2 CH 2 -C(=O)-);

[0310] A disulfide bond is formed between the 2nd and 12th cysteines;

[0311] wherein the C-terminus of SEQ ID NO: 20 is amidated.

[0312] In a specific embodiment, the present invention provides a polypeptide having the amino acid sequence shown in SEQ ID NO: 28 as follows; or having at least 95%, preferably at least 94%, at least 93%, at least 92%, at least 91%, at least 90%, at least 89%, at least 87%, at least 86%, at least 85%, at least 84%, at least 83%, at least 82%, at least 81%, at least 80%, at least 79%, at least 78%, at least 77%, at least 76%, at least 75% sequence identity with the amino acid sequence shown in SEQ ID NO: 28; or having up to 3, preferably up to 2 or up to 1 amino acid sequence substitutions, deletions or additions compared to SEQ ID NO: 28:

[0313] {D-Tyr}-ICV-{Trp(Me)}-QDW-(Sar)-AHRCI, (SEQ ID NO: 28); wherein D-Tyr represents D-tyrosine, Trp(Me) represents N-methyltryptophan, Sar represents sarcosine, and a disulfide bond is formed between Cys3-Cys13.

[0314] In a specific embodiment, the present invention provides a polypeptide having the amino acid sequence shown in SEQ ID NO: 29 as follows; or having at least 95%, preferably at least 94%, at least 93%, at least 92%, at least 91%, at least 90%, at least 89%, at least 87%, at least 86%, at least 85%, at least 84%, at least 83%, at least 82%, at least 81%, at least 80%, at least 79%, at least 78%, at least 77%, at least 76%, at least 75% sequence identity with the amino acid sequence shown in SEQ ID NO: 29; or having up to 3, preferably up to 2 or up to 1 amino acid sequence substitutions, deletions or additions compared to SEQ ID NO: 29:

[0315] {D-Tyr}-ICV-{Trp(Me)}-QDW-(Sar)-AHRCIGRKKRRQRRRPQ (SEQ ID NO: 29);

[0316] Wherein D-Tyr represents D-tyrosine, Trp(Me) represents N-methyltryptophan, Sar represents sarcosine, and a disulfide bond is formed between Cys3-Cys13, wherein the C-terminus of SEQ ID NO: 29 is amidated.

[0317] In a specific embodiment, the present invention provides a polypeptide having the amino acid sequence shown in SEQ ID NO:30; or having at least 95%, preferably at least 94%, at least 93%, at least 92%, at least 91%, at least 90%, at least 89%, at least 87%, at least 86%, at least 85%, at least 84%, at least 83%, at least 82%, at least 81%, at least 80%, at least 79%, at least 78%, at least 77%, at least 76%, at least 75% sequence identity with the amino acid sequence shown in SEQ ID NO:30; or having at most 3, preferably at most 2 or at most 1 amino acid sequence substitutions, deletions or additions compared to SEQ ID NO:30:

[0318] {D-Tyr}-ICV-{Trp(Me)}-QDW-(Sar)-AHRCI-(AEEA)-GRKKRRQRRRPQ (SEQ ID NO: 30);

[0319] Where D-Tyr represents D-tyrosine, Trp(Me) represents N-methyltryptophan, Sar represents sarcosine, and AEEA represents 8-amino-3,6-dioxaoctanoyl (-NH-CH 2 CH 2 OCH 2 CH 2 -C(=O)-), and a disulfide bond is formed between Cys3-Cys13, wherein the C-terminus of SEQ ID NO: 30 is amidated.

[0320] In a specific embodiment, the present invention provides a polypeptide having the amino acid sequence shown in SEQ ID NO:31 as follows; or having at least 95%, preferably at least 94%, at least 93%, at least 92%, at least 91%, at least 90%, at least 89%, at least 87%, at least 86%, at least 85%, at least 84%, at least 83%, at least 82%, at least 81%, at least 80%, at least 79%, at least 78%, at least 77%, at least 76%, at least 75% sequence identity with the amino acid sequence shown in SEQ ID NO:31; or having up to 3, preferably up to 2 or up to 1 amino acid sequence substitutions, deletions or additions compared to SEQ ID NO:31:

[0321] {D-Tyr}-ICV-{Trp(Me)}-QDW-(Sar)-AHRCIRQIKIWFQNRR MKWKK (SEQ ID NO: 31);

[0322] Wherein D-Tyr represents D-tyrosine, Trp(Me) represents N-methyltryptophan, Sar represents sarcosine, and a disulfide bond is formed between Cys3-Cys13, wherein the C-terminus of SEQ ID NO: 31 is amidated.

[0323] In a specific embodiment, the present invention provides a polypeptide having the amino acid sequence shown in SEQ ID NO:32 as follows; or having at least 95%, preferably at least 94%, at least 93%, at least 92%, at least 91%, at least 90%, at least 89%, at least 87%, at least 86%, at least 85%, at least 84%, at least 83%, at least 82%, at least 81%, at least 80%, at least 79%, at least 78%, at least 77%, at least 76%, at least 75% sequence identity with the amino acid sequence shown in SEQ ID NO:32; or having up to 3, preferably up to 2 or up to 1 amino acid sequence substitutions, deletions or additions compared to SEQ ID NO:32:

[0324] {D-Tyr}-ICV-{Trp(Me)}-QDW-(Sar)-AHRCI-(AEEA)-RQIKIWFQNRRMKWKK(SEQ IDNO:32);

[0325] Where D-Tyr represents D-tyrosine, Trp(Me) represents N-methyltryptophan, Sar represents sarcosine, and AEEA represents 8-amino-3,6-dioxaoctanoyl (-NH-CH 2 CH 2 OCH 2 CH 2 -C(=O)-), and a disulfide bond is formed between Cys3-Cys13, wherein the C-terminus of SEQ ID NO: 32 is amidated.

[0326] In a specific embodiment, the present invention provides a polypeptide having the amino acid sequence shown in SEQ ID NO:33 as follows; or having at least 95%, preferably at least 94%, at least 93%, at least 92%, at least 91%, at least 90%, at least 89%, at least 87%, at least 86%, at least 85%, at least 84%, at least 83%, at least 82%, at least 81%, at least 80%, at least 79%, at least 78%, at least 77%, at least 76%, at least 75% sequence identity with the amino acid sequence shown in SEQ ID NO:33; or having at most 3, preferably at most 2 or at most 1 amino acid sequence substitutions, deletions or additions compared to SEQ ID NO:33:

[0327] {D-Tyr}-ICV-{Trp(Me)}-QDW-(Sar)-AHRCI-(AEEA)-GRKKRRQRRRPQ(SEQ ID NO:33)

[0328] Where D-Tyr represents D-tyrosine, Trp(Me) represents N-methyltryptophan, Sar represents sarcosine, and AEEA represents 8-amino-3,6-dioxaoctanoyl (-NH-CH 2 CH 2 OCH 2 CH 2 -C(=O)-), and a disulfide bond is formed between Cys3-Cys13.

[0329] In a specific embodiment, the present invention provides a polypeptide having the amino acid sequence shown in SEQ ID NO:34 as follows; or having at least 95%, preferably at least 94%, at least 93%, at least 92%, at least 91%, at least 90%, at least 89%, at least 87%, at least 86%, at least 85%, at least 84%, at least 83%, at least 82%, at least 81%, at least 80%, at least 79%, at least 78%, at least 77%, at least 76%, at least 75% sequence identity with the amino acid sequence shown in SEQ ID NO:34; or having at most 3, preferably at most 2 or at most 1 amino acid sequence substitutions, deletions or additions compared to SEQ ID NO:34:

[0330] {D-Tyr}-ICV-{Trp(Me)}-QDW-(Sar)-AHRCI-(AEEA)-YGRKKRRQRRR(SEQ ID NO:34)

[0331] Where D-Tyr represents D-tyrosine, Trp(Me) represents N-methyltryptophan, Sar represents sarcosine, and AEEA represents 8-amino-3,6-dioxaoctanoyl (-NH-CH 2 CH 2 OCH 2 CH 2 -C(=O)-), and a disulfide bond is formed between Cys3-Cys13.

[0332] In a specific embodiment, the present invention provides a polypeptide having the amino acid sequence shown in SEQ ID NO:35 as follows; or having at least 95%, preferably at least 94%, at least 93%, at least 92%, at least 91%, at least 90%, at least 89%, at least 87%, at least 86%, at least 85%, at least 84%, at least 83%, at least 82%, at least 81%, at least 80%, at least 79%, at least 78%, at least 77%, at least 76%, at least 75% sequence identity with the amino acid sequence shown in SEQ ID NO:35; or having at most 3, preferably at most 2 or at most 1 amino acid sequence substitutions, deletions or additions compared to SEQ ID NO:35:

[0333] {D-Tyr}-ICV-{Trp(Me)}-QDW-(Sar)-AHRCI-(AEEA)-GYGRKKRRQRRR(SEQ ID NO:35)

[0334] Where D-Tyr represents D-tyrosine, Trp(Me) represents N-methyltryptophan, Sar represents sarcosine, and AEEA represents 8-amino-3,6-dioxaoctanoyl (-NH-CH 2 CH 2 OCH 2 CH 2 -C(=O)-), and a disulfide bond is formed between Cys3-Cys13; wherein the C-terminus of SEQ ID NO: 35 is amidated.

[0335] In a specific embodiment, the present invention provides a polypeptide having the amino acid sequence shown in SEQ ID NO:36 as follows; or having at least 95%, preferably at least 94%, at least 93%, at least 92%, at least 91%, at least 90%, at least 89%, at least 87%, at least 86%, at least 85%, at least 84%, at least 83%, at least 82%, at least 81%, at least 80%, at least 79%, at least 78%, at least 77%, at least 76%, at least 75% sequence identity with the amino acid sequence shown in SEQ ID NO:36; or having at most 3, preferably at most 2 or at most 1 amino acid sequence substitutions, deletions or additions compared to SEQ ID NO:36:

[0336] {D-Tyr}-ICV-{Trp(Me)}-QDW-(Sar)-AHRCI-(AEEA)-GGYGRKKRRQRRR(SEQ ID NO:36)

[0337] Where D-Tyr represents D-tyrosine, Trp(Me) represents N-methyltryptophan, Sar represents sarcosine, and AEEA represents 8-amino-3,6-dioxaoctanoyl (-NH-CH 2 CH 2 OCH 2 CH 2 -C(=O)-), and a disulfide bond is formed between Cys3-Cys13; wherein the C-terminus of SEQ ID NO: 36 is amidated.

[0338] In a specific embodiment, the present invention provides a polypeptide having the amino acid sequence shown in SEQ ID NO:37 as follows; or having at least 95%, preferably at least 94%, at least 93%, at least 92%, at least 91%, at least 90%, at least 89%, at least 87%, at least 86%, at least 85%, at least 84%, at least 83%, at least 82%, at least 81%, at least 80%, at least 79%, at least 78%, at least 77%, at least 76%, at least 75% sequence identity with the amino acid sequence shown in SEQ ID NO:37; or having up to 3, preferably up to 2 or up to 1 amino acid sequence substitutions, deletions or additions compared to SEQ ID NO:37:

[0339] {D-Tyr}-ICV-{Trp(Me)}-QDW-(Sar)-AHRCI-(AEEA)-GGGYGRKKRRQRRR(SEQ IDNO:37)

[0340] Where D-Tyr represents D-tyrosine, Trp(Me) represents N-methyltryptophan, Sar represents sarcosine, and AEEA represents 8-amino-3,6-dioxaoctanoyl (-NH-CH 2 CH 2 OCH 2 CH 2 -C(=O)-), and a disulfide bond is formed between Cys3-Cys13; wherein the C-terminus of SEQ ID NO: 37 is amidated.

[0341] In a specific embodiment, the present invention provides a polypeptide having the amino acid sequence shown in SEQ ID NO:38 as follows; or having at least 95%, preferably at least 94%, at least 93%, at least 92%, at least 91%, at least 90%, at least 89%, at least 87%, at least 86%, at least 85%, at least 84%, at least 83%, at least 82%, at least 81%, at least 80%, at least 79%, at least 78%, at least 77%, at least 76%, at least 75% sequence identity with the amino acid sequence shown in SEQ ID NO:38; or having at most 3, preferably at most 2 or at most 1 amino acid sequence substitutions, deletions or additions compared to SEQ ID NO:38:

[0342] {ACI}-CV-{Trp(Me)}-QDWGAHRCT (SEQ ID NO: 38);

[0343] ACI represents acetylated isoleucine, Trp(Me) represents N-methyltryptophan, and a disulfide bond is formed between Cys2 and Cys12.

[0344] In a specific embodiment, the present invention provides a polypeptide having the amino acid sequence shown in SEQ ID NO:39 as follows; or having at least 95%, preferably at least 94%, at least 93%, at least 92%, at least 91%, at least 90%, at least 89%, at least 87%, at least 86%, at least 85%, at least 84%, at least 83%, at least 82%, at least 81%, at least 80%, at least 79%, at least 78%, at least 77%, at least 76%, at least 75% sequence identity with the amino acid sequence shown in SEQ ID NO:39; or having up to 3, preferably up to 2 or up to 1 amino acid sequence substitutions, deletions or additions compared to SEQ ID NO:39:

[0345] {ACI}-CV-{Trp(Me)}-QDWGAHRCT GRKKRRQRRRPQ(SEQ ID NO:39)

[0346] Wherein ACI represents acetylated isoleucine, Trp(Me) represents N-methyltryptophan, and a disulfide bond is formed between Cys2-Cys12, wherein the C-terminus of SEQ ID NO: 39 is amidated.

[0347] In a specific embodiment, the present invention provides a polypeptide having the amino acid sequence shown in SEQ ID NO:40 as follows; or having at least 95%, preferably at least 94%, at least 93%, at least 92%, at least 91%, at least 90%, at least 89%, at least 87%, at least 86%, at least 85%, at least 84%, at least 83%, at least 82%, at least 81%, at least 80%, at least 79%, at least 78%, at least 77%, at least 76%, at least 75% sequence identity with the amino acid sequence shown in SEQ ID NO:40; or having up to 3, preferably up to 2 or up to 1 amino acid sequence substitutions, deletions or additions compared to SEQ ID NO:40:

[0348] {ACI}-CV-{Trp(Me)}-QDWGAHRCT-(AEEA)-GRKKRRQRRRPQ(SEQ ID NO:40)

[0349] ACI represents acetylated isoleucine, Trp(Me) represents N-methyltryptophan, and a disulfide bond is formed between Cys2 and Cys12. AEEA represents 8-amino-3,6-dioxaoctanoyl (-NH-CH 2 CH 2 OCH 2 CH 2 -C(=O)-), wherein the C-terminus of SEQ ID NO: 40 is amidated.

[0350] In a specific embodiment, the present invention provides a polypeptide having the amino acid sequence shown in SEQ ID NO:41 as follows; or having at least 95%, preferably at least 94%, at least 93%, at least 92%, at least 91%, at least 90%, at least 89%, at least 87%, at least 86%, at least 85%, at least 84%, at least 83%, at least 82%, at least 81%, at least 80%, at least 79%, at least 78%, at least 77%, at least 76%, at least 75% sequence identity with the amino acid sequence shown in SEQ ID NO:41; or having up to 3, preferably up to 2 or up to 1 amino acid sequence substitutions, deletions or additions compared to SEQ ID NO:41:

[0351] {ACI}-CV-{Trp(Me)}-QDWGAHRCTRQIKIWFQNRRMKWKK(SEQ ID NO:41)

[0352] Wherein ACI represents acetylated isoleucine, Trp(Me) represents N-methyltryptophan, and a disulfide bond is formed between Cys2-Cys12, wherein the C-terminus of SEQ ID NO:41 is amidated.

[0353] In a specific embodiment, the present invention provides a polypeptide having the amino acid sequence shown in SEQ ID NO:42 as follows; or having at least 95%, preferably at least 94%, at least 93%, at least 92%, at least 91%, at least 90%, at least 89%, at least 87%, at least 86%, at least 85%, at least 84%, at least 83%, at least 82%, at least 81%, at least 80%, at least 79%, at least 78%, at least 77%, at least 76%, at least 75% sequence identity with the amino acid sequence shown in SEQ ID NO:42; or having up to 3, preferably up to 2 or up to 1 amino acid sequence substitutions, deletions or additions compared to SEQ ID NO:42:

[0354] {ACI}-CV-{Trp(Me)}-QDWGAHRCT-(AEEA)-RQIKIWFQNRRMKWKK(SEQ ID NO:42)

[0355] ACI represents acetylated isoleucine, Trp(Me) represents N-methyltryptophan, and a disulfide bond is formed between Cys2 and Cys12. AEEA represents 8-amino-3,6-dioxaoctanoyl (-NH-CH 2 CH 2 OCH 2 CH 2 -C(=O)-), wherein the C-terminus of SEQ ID NO: 42 is amidated.

[0356] In a specific embodiment, the present invention provides a polypeptide having the amino acid sequence shown in SEQ ID NO:43 as follows; or having at least 95%, preferably at least 94%, at least 93%, at least 92%, at least 91%, at least 90%, at least 89%, at least 87%, at least 86%, at least 85%, at least 84%, at least 83%, at least 82%, at least 81%, at least 80%, at least 79%, at least 78%, at least 77%, at least 76%, at least 75% sequence identity with the amino acid sequence shown in SEQ ID NO:43; or having up to 3, preferably up to 2 or up to 1 amino acid sequence substitutions, deletions or additions compared to SEQ ID NO:43:

[0357] {ACI}-CV-{Trp(Me)}-QDWGAHRCT-(AEEA)-GRKKRRQRRRPQ(SEQ ID NO:43)

[0358] ACI represents acetylated isoleucine, Trp(Me) represents N-methyltryptophan, and a disulfide bond is formed between Cys2 and Cys12. AEEA represents 8-amino-3,6-dioxaoctanoyl (-NH-CH 2 CH 2 OCH 2 CH 2 -C(=O)-).

[0359] In a specific embodiment, the present invention provides a polypeptide having the amino acid sequence shown in SEQ ID NO:44 as follows; or having at least 95%, preferably at least 94%, at least 93%, at least 92%, at least 91%, at least 90%, at least 89%, at least 87%, at least 86%, at least 85%, at least 84%, at least 83%, at least 82%, at least 81%, at least 80%, at least 79%, at least 78%, at least 77%, at least 76%, at least 75% sequence identity with the amino acid sequence shown in SEQ ID NO:44; or having up to 3, preferably up to 2 or up to 1 amino acid sequence substitutions, deletions or additions compared to SEQ ID NO:44:

[0360] {ACI}-CV-{Trp(Me)}-QDWGAHRCT-(AEEA)-YGRKKRRQRRR(SEQ ID NO:44)

[0361] ACI represents acetylated isoleucine, Trp(Me) represents N-methyltryptophan, and a disulfide bond is formed between Cys2 and Cys12. AEEA represents 8-amino-3,6-dioxaoctanoyl (-NH-CH 2 CH 2 OCH 2 CH 2 -C(=O)-), wherein the C-terminus of SEQ ID NO: 44 is amidated.

[0362] In a specific embodiment, the present invention provides a polypeptide having the amino acid sequence shown in SEQ ID NO:45 as follows; or having at least 95%, preferably at least 94%, at least 93%, at least 92%, at least 91%, at least 90%, at least 89%, at least 87%, at least 86%, at least 85%, at least 84%, at least 83%, at least 82%, at least 81%, at least 80%, at least 79%, at least 78%, at least 77%, at least 76%, at least 75% sequence identity with the amino acid sequence shown in SEQ ID NO:45; or having up to 3, preferably up to 2 or up to 1 amino acid sequence substitutions, deletions or additions compared to SEQ ID NO:45:

[0363] {ACI}-CV-{Trp(Me)}-QDWGAHRCT-(AEEA)-GYGRKKRRQRRR(SEQ ID NO:45)

[0364] ACI represents acetylated isoleucine, Trp(Me) represents N-methyltryptophan, and a disulfide bond is formed between Cys2 and Cys12. AEEA represents 8-amino-3,6-dioxaoctanoyl (-NH-CH 2 CH 2 OCH 2 CH 2 -C(=O)-), wherein the C-terminus of SEQ ID NO: 45 is amidated.

[0365] In a specific embodiment, the present invention provides a polypeptide having the amino acid sequence shown in SEQ ID NO:46 as follows; or having at least 95%, preferably at least 94%, at least 93%, at least 92%, at least 91%, at least 90%, at least 89%, at least 87%, at least 86%, at least 85%, at least 84%, at least 83%, at least 82%, at least 81%, at least 80%, at least 79%, at least 78%, at least 77%, at least 76%, at least 75% sequence identity with the amino acid sequence shown in SEQ ID NO:46; or having up to 3, preferably up to 2 or up to 1 amino acid sequence substitutions, deletions or additions compared to SEQ ID NO:46:

[0366] {ACI}-CV-{Trp(Me)}-QDWGAHRCT-(AEEA)-GGYGRKKRRQRRR(SEQ ID NO:46)

[0367] ACI represents acetylated isoleucine, Trp(Me) represents N-methyltryptophan, and a disulfide bond is formed between Cys2 and Cys12. AEEA represents 8-amino-3,6-dioxaoctanoyl (-NH-CH 2 CH 2 OCH 2 CH 2 -C(=O)-), wherein the C-terminus of SEQ ID NO: 46 is amidated.

[0368] In a specific embodiment, the present invention provides a polypeptide having the amino acid sequence shown in SEQ ID NO:47 as follows; or having at least 95%, preferably at least 94%, at least 93%, at least 92%, at least 91%, at least 90%, at least 89%, at least 87%, at least 86%, at least 85%, at least 84%, at least 83%, at least 82%, at least 81%, at least 80%, at least 79%, at least 78%, at least 77%, at least 76%, at least 75% sequence identity with the amino acid sequence shown in SEQ ID NO:47; or having up to 3, preferably up to 2 or up to 1 amino acid sequence substitutions, deletions or additions compared to SEQ ID NO:47:

[0369] {ACI}-CV-{Trp(Me)}-QDWGAHRCT-(AEEA)-GGGYGRKKRRQRRR(SEQ ID NO:47)

[0370] ACI represents acetylated isoleucine, Trp(Me) represents N-methyltryptophan, and a disulfide bond is formed between Cys2 and Cys12. AEEA represents 8-amino-3,6-dioxaoctanoyl (-NH-CH 2 CH 2 OCH 2 CH 2 -C(=O)-), wherein the C-terminus of SEQ ID NO: 47 is amidated.

[0371] In a specific embodiment, the present invention provides a group of polypeptides capable of passing through a cell membrane or a tissue membrane, comprising any of the following amino acid sequences:

[0372] GRKKRRQRRRPQ (SEQ ID NO: 21);

[0373] RQIKIWFQNRRMKWKK (SEQ ID NO: 22);

[0374] GRKKRRQRRRPQ (SEQ ID NO: 23);

[0375] YGRKKRRQRRR (SEQ ID NO: 24);

[0376] GYGRKKRRQRRR (SEQ ID NO: 25);

[0377] GGYGRKKRRQRRR (SEQ ID NO: 26); or

[0378] GGGYGRKKRRQRRR (SEQ ID NO: 27);

[0379] Optionally, the polypeptide shown in any one of SEQ ID NO: 21 to SEQ ID NO: 27 has or does not have C-terminal amidation.

[0380] In a specific embodiment, the polypeptide provided by the present invention may have an acetylation modification at the N-terminus and / or an amidation modification at the C-terminus.

[0381] Acetylation of the N-terminus and / or amidation of the C-terminus has several advantages:

[0382] It can eliminate the terminal charge of the polypeptide, making it closer to the parent protein and enhancing its ability to enter the cell; and

[0383] It can enhance the stability of polypeptides and improve their resistance to exopeptidases such as proteases, endoplasmic reticulum and synthetases.

[0384] The present invention provides the transmembrane functional polypeptide that penetrates the cell membrane or body tissue barrier, or is used to connect with a functional polypeptide to prepare a complex that can penetrate the cell membrane or body tissue barrier, or is used to prepare a pharmaceutical composition that can penetrate the cell membrane or body tissue barrier.

[0385] In a preferred embodiment, the functional polypeptides include (but are not limited to): anti-angiogenesis polypeptides, the functional molecules of which are C3 complement inhibitors.

[0386] In a preferred example, the body tissue barrier is an eye barrier, and preferably the eye barrier includes: eye tissue barrier (the various layers of tissues forming the eyeball wall and the cell connections between the various layers of tissues, etc.), tear barrier, blood-eye barrier (blood-aqueous humor barrier, blood-retinal barrier). In a preferred example, the pharmaceutical composition is a pharmaceutical composition for preventing or treating eye diseases, and its dosage forms include: eye drop dosage forms, subconjunctival injection dosage forms, periocular injection dosage forms, subretinal injection dosage forms, and intraocular administration (such as intravitreal injection) dosage forms.

[0387] In preferred embodiments, the eye diseases include (but are not limited to): age-related macular degeneration (AMD) and diabetic macular edema (DME), inflammatory eye diseases, and optic neuropathy.

[0388] In some embodiments of the present invention, the ocular microangiopathy-related disorder is selected from diabetic retinopathy (DR), diabetic macular edema (DME), retinal vein occlusion (RVO), central retinal vein occlusion (CRVO), macular degeneration, retinopathy of prematurity (ROP), neovascular glaucoma, retinitis pigmentosa (RP), proliferative and non-proliferative retinopathy, retinal angiomatous proliferation, macular telangiectasia, ischemic retinopathy, iris neovascularization, intraocular neovascularization, corneal neovascularization, retinal neovascularization, polypoidal choroidal vasculopathy (PCV), choroidal neovascularization, retinal degeneration, diabetic complications of retinal vascular permeability associated with diabetic retinopathy and diabetic macular edema; in particular, selected from diabetic retinopathy (DR), diabetic macular edema (DME), retinal vein occlusion (RVO), central retinal vein occlusion (CRVO) and wet age-related macular degeneration (wet AMD).

[0389] In specific embodiments, the present invention provides pharmaceutical compositions comprising a polypeptide or fusion polypeptide described herein, and one or more pharmaceutically acceptable carriers or excipients.

[0390] The pharmaceutical composition of the present invention may be in liquid form. On the one hand, the carrier is a pharmaceutically acceptable carrier in the composition that may contain a liquid, such as water, phosphate buffer, Ringer solution, physiological saline, balanced salt solution, glycerol or sorbitol, etc. In addition, these carriers may also contain auxiliary substances, such as lubricants, glidants, wetting agents or emulsifiers, pH buffer substances and stabilizers, such as albumin, etc.

[0391] The pharmaceutical composition can be administered by eye drops, subconjunctival injection, periocular injection, subretinal injection, or intraocular injection.

[0392] The eye administration refers to the administration of eye drops, eye gels or eye ointments on the ocular surface in a non-invasive manner.

[0393] The compounds and pharmaceutical compositions of the present invention may be further combined with one or more, preferably one, additional therapeutic agent. The compounds of the present invention may be combined with one or more additional therapeutic agents selected from the group consisting of: anti-VEGF monoclonal or dual antibodies, antidiabetic agents, agents for treating overweight and / or obesity; agents for treating hypertension, pulmonary hypertension, heart failure and / or atherosclerosis, and agents for treating eye diseases.

[0394] The drug that can be combined with other drugs for treating type II diabetes is selected from GK agonists, GLP-1 receptor agonists, exenatide, liraglutide, semaglutide, dulaglutide, loxenatide, albiglutide; α-glucosidase inhibitors, voglibose, acarbose; SGLT-2 inhibitors, canagliflozin, dapagliflozin, empagliflozin, ipragliflozin, rupagliflozin Luseogliflozin and Tofogliflozin; DPP-4 inhibitors, one or more of sitagliptin, vildagliptin, saxagliptin, alogliptin, linagliptin, gemigliptin and teneligliptin and metformin. The pharmaceutical composition can be used to prevent one or more metabolic disorders selected from the following, slow down the progression of the metabolic disorder, or treat the metabolic disorder: type I diabetes, type II diabetes, impaired glucose tolerance, hyperglycemia, postprandial hyperglycemia, fasting blood glucose abnormalities, overweight, obesity, hypertension, insulin resistance and / or metabolic syndrome; or improve blood sugar control and / or reduce fasting plasma glucose; or prevent, slow, delay or reverse diabetic complications.

[0395] It can be combined with other drugs to treat hypertension, angiotensin II receptor blockers (ARB), angiotensin converting enzyme inhibitors (ACEI), calcium channel blockers (CCB), beta blockers, diuretics, alpha blockers, etc. Selected from felodipine sustained-release tablets, nifedipine, nitrendipine, nisoldipine, nicardipine, lacidipine, lercanidipine, amlodipine, levamlodipine, verapamil, diltiazem, captopril, enalapril, benazepril, lislopril, ramipril, fosinopril, cilazapril, perindopril, quinapril, imidapril, zofenopril, losartan, valsartan, irbesartan, irbesartan, candesartan cilexetil, telmisartan, olmesartan medoxomil, etc.

[0396] Drugs that can be combined with other agents to treat heart failure and / or atherosclerosis are selected from mevastatin, lovastatin, pravastatin, simvastatin, fluvastatin, cholestyramine, colestipol, aspirin, clopidogrel, cilostazol, ω-3 polyunsaturated fatty acids, ω-6 monounsaturated fatty acids, heparin, heparan sulfate, chondroitin sulfate, dextran sulfate, finerenone, proprotein convertase subtilisin / kexin type 9 (PCSK9) type inhibitors evolocumab (Rebai'an, Evolocumab), alirocumab (Polida, Alirocumab), Inclisiran (Leqvio), etc.

[0397] It can be combined with other drugs to treat pulmonary hypertension, selected from bosentan, riociguat, macitentan, selexipag and the like.

[0398] Other therapeutic agents that can be combined with it for the treatment of ocular diseases include intravitreally administered corticosteroids, intravitreally administered anti-VEGF therapeutics, anti-Ang2 inhibitors, dual anti-VEGF / anti-Ang2 inhibitors, supplementation inhibitors (supplementation factor 3, 5, B and D inhibitors, etc.), anti-PDGF, dual anti-VEGF / anti-PDGF, VAP-1 inhibitors, bradykinin receptor 1 (BK1) antagonists, Tie-2 agonists, CCR-2 antagonists, etc.

[0399] The drug that can be combined with other drugs, such as monoclonal or dual anti-VEGF antibodies, is selected from aflibercept (Eylea), bevacizumab (Avastin), ranibizumab (Lucentis), pegaptanib (Macugen), and faricimab (Vabysmo).

[0400] The compounds provided by the present invention can be prepared using the specific synthesis schemes described below from readily available starting materials, which can be synthesized using or according to methods known in the art, or can be purchased commercially. The experimental methods in the following examples where specific conditions are not indicated can be determined by conventional methods and conditions by those skilled in the art through conventional optimization procedures.

[0401] Preparation Example 1. Preparation, modification and identification of C3 complement inhibitory polypeptide

[0402] Specifically, the polypeptide provided by the present invention is prepared by using a twelve-channel semi-automatic polypeptide synthesizer to synthesize a fusion polypeptide. The specific operation process is as follows:

[0403] Put Rink Amide-MBHA Resin into a reaction tube, add solvent DCM, and shake for 30-40 minutes. Add 20% piperidine DMF solution. Filter out the solvent through the sand core and wash it clean. Then add 3 times molar excess of Fmoc protected amino acid (the first amino acid at the C-terminus), add DMF to dissolve, 3 times excess of condensing agent HBTU or HATU, and then add 10 times molar excess of DIEA, and shake for 60-80 minutes. Block with pyridine and acetic anhydride. Remove DMF, add 20% piperidine DMF solution (15ml / g), rinse for 5-10 minutes, remove and add 20% piperidine DMF solution. Drain the piperidine solution, take a dozen resins, wash three times with ethanol, add one drop each of ninhydrin, KCN, and phenol solution, heat at 105℃-110℃ for 5-10 minutes, and turn dark blue for positive reaction. Rinse twice with DMF, twice with methanol, and twice with DMF. Add 3 times molar excess of Fmoc protected amino acid, 3 times molar excess of HBTU, and then add 10 times molar excess of DIEA, and finally add DMF to dissolve, and shake for 45-50 minutes. Take a dozen resins, wash three times with ethanol, add one drop of ninhydrin, pyridine, and phenol solution, heat at 105℃-110℃ for 5 minutes, and colorless is a negative reaction. Rinse once with DMF, rinse twice with methanol, and rinse twice with DMF. Repeat the above operation, connect from right to left in sequence, until the Fmoc protecting group of the last amino acid is removed, and then acetylate and cap. Wash the resin according to the following method, DMF (10ml / g) twice, DCM (10ml / g) three times, methanol (10ml / g) four times, and drain for 10 minutes. Prepare cutting solution TFA 95%; water 2%; EDT 2%; TIS1% Cutting time: 180-200min. The lysate was blown dry as much as possible with nitrogen, ether was precipitated, the supernatant was removed by centrifugation, the precipitate was washed with ether six times, and then evaporated at room temperature. After the dried crude product was completely dissolved, the disulfide bonds were oxidized by DMSO oxidation method, and Ellman reagent was used for monitoring during the process until the oxidation was complete. Take a small amount of crude product and dissolve it in H2O / ACN. Take a small amount of sample and analyze it on the HPLC analyzer to determine the peak time corresponding to the target peak. Use C18 reverse phase chromatography preparation system: Wavelength: 220nm; Flow Rate: 15ml / min; Inj.Vol: 20mL Column Temp: 25℃; Buffer A: 0.1% TFA in water Buffer B: 0.1% TFA in Acetonitrile; collect the target peak solution. Use a 1.5ml centrifuge tube to take a small amount of the target peak solution for mass spectrometry confirmation and purity detection. The qualified target peak solution was freeze-dried to obtain the finished product. The sample was re-dissolved in H2O / ACN and salted on the HPLC analyzer. The solution after the salt conversion treatment is freeze-dried for a second time to obtain a salt-type finished product.

[0404] Exemplary polypeptides were synthesized as follows

[0405] {D-Tyr}-ICV-{Trp(Me)}-QDW-(Sar)-AHRCI, (SEQ ID NO: 28); wherein D-Tyr represents D-tyrosine, Trp(Me) represents N-methyltryptophan, Sar represents sarcosine, and a disulfide bond is formed between Cys3-Cys13.

[0406] Synthesis method: Fmoc solid phase synthesis.

[0407] Synthetic raw materials and related reagents:

[0408] 1. Protect amino acid raw materials

[0409] Fmoc-Ile-OH, Fmoc-Cys(Trt)-OH, Fmoc-Val-OH, Fmoc-Trp(1-Me)-OH, Fmoc-Gln(Trt)-OH, Fmoc-Asp(otbu) -OH, Fmoc-Trp(Boc)-OH, Fmoc-Gly-OH, Fmoc-Ala-OH, Fmoc-His(Trt)-OH, Fmoc-Arg(pbf)-OH, Fmoc-Thr(tb u)-OH

[0410] 2. Condensation reagents HBTU, DIEA

[0411] 3. Solvent

[0412] DMF, DCM, methanol, acetonitrile

[0413] 4. Resin

[0414] Rink Amide-MBHA Resin with a substitution degree of 0.38mmol / g

[0415] 5. Deprotection reagent piperidine

[0416] 6. Detection reagents:

[0417] Phenol reagent, pyridine reagent, ninhydrin reagent

[0418] 7. Cutting Reagents

[0419] TFA, TIS, EDT, anhydrous ether

[0420] 8. Equipment

[0421] a) Precision electronic balance equipment:

[0422] b) Twelve-channel semi-automatic peptide synthesizer

[0423] c) High performance liquid chromatography

[0424] d) freeze dryer,

[0425] e) Centrifuge.

[0426] Synthesis process:

[0427] Resin swelling

[0428] Place Rink Amide-MBHA Resin in a reaction tube, add DCM (15 ml / g), and shake for 30 min.

[0429] First amino acid

[0430] Add 20% piperidine DMF solution (15ml / g), 15min. Filter out the solvent through a sand core and wash it clean. Add 3 times molar excess of Fmoc protected amino acid (first amino acid at the C-terminus), add DMF to dissolve, 3 times excess of HBTU, and then add 10 times molar excess of DIEA, shake for 60min. Block with pyridine and acetic anhydride.

[0431] Deprotection

[0432] Remove DMF, add 20% piperidine DMF solution (15 ml / g) for 5 min, remove and add 20% piperidine DMF solution (15 ml / g) for 15 min.

[0433] Test

[0434] Drain off the piperidine solution, take a dozen resin pellets, wash them three times with ethanol, add one drop each of ninhydrin, KCN, and phenol solution, heat at 105℃-110℃ for 5 minutes, and a dark blue color indicates a positive reaction.

[0435] rinse

[0436] DMF (10 ml / g) twice, methanol (10 ml / g) twice, DMF (10 ml / g) twice

[0437] condensation

[0438] A 3-fold molar excess of Fmoc-protected amino acid, a 3-fold molar excess of HBTU, and a 10-fold molar excess of DIEA were added, and finally DMF was added to dissolve and shaken for 45 min.

[0439] Detection

[0440] Take a dozen resin particles, wash them three times with ethanol, add one drop each of ninhydrin, pyridine and phenol solution, heat at 105℃-110℃ for 5 minutes, and colorless is a negative reaction.

[0441] rinse

[0442] DMF (10 ml / g) once, methanol (10 ml / g) twice, DMF (10 ml / g) twice

[0443] Repeat steps 3 to 8, connecting from right to left, until the Fmoc protecting group of the last amino acid is removed, and then acetylation is performed to cap the end.

[0444] Wash the resin and drain it as follows:

[0445] DMF (10 ml / g) twice, DCM (10 ml / g) three times, methanol (10 ml / g) four times, and pump dry for 10 min.

[0446] Cutting

[0447] Prepare cutting solution (10 / g): TFA 95%; water 2%; EDT 2%; TIS 1% Cutting time: 180 min

[0448] Blow dry wash

[0449] The lysate was blown dry with nitrogen as much as possible, ether was precipitated, the supernatant was removed by centrifugation, the precipitate was washed with ether six times, and then evaporated to dryness at room temperature.

[0450] Oxidation

[0451] After the dried crude product is completely dissolved, the disulfide bonds are oxidized using the DMSO oxidation method, and the Ellman reagent is used for monitoring during the process until the oxidation is complete.

[0452] Purification preparation.

[0453] a) Take a small amount of crude product and dissolve it in H2O / ACN.

[0454] b) Take a small amount of sample and analyze it on HPLC to determine the peak time of the target peak.

[0455] c) Using C18 reverse phase chromatography system: Wavelength: 220nm; Flow Rate: 15ml / min; Inj.Vol: 20mL Column Temp: 25℃

[0456] d) Buffer A: 0.1% TFA in water Buffer B: 0.1% TFA in Acetonitrile; collect the target peak solution.

[0457] e) Take a small amount of the target peak solution in a 1.5 ml centrifuge tube for mass spectrometry confirmation and purity testing.

[0458] The qualified target peak solution is freeze-dried to obtain the finished product.

[0459] The sample was redissolved in H2O / ACN and subjected to salt conversion on an HPLC analyzer.

[0460] The solution after the salt conversion treatment is freeze-dried for a second time to obtain a finished product.

[0461] Identification

[0462] Take a small amount of the finished peptide for molecular weight identification by MS and purity identification by HPLC analysis.

[0463] The following peptides were prepared using the same method as above:

[0464] {D-Tyr}-ICV-{Trp(Me)}-QDW-(Sar)-AHRCIGRKKRRQRRRPQ (SEQ ID NO: 29);

[0465] wherein D-Tyr represents D-tyrosine, Trp(Me) represents N-methyltryptophan, Sar represents sarcosine, and a disulfide bond is formed between Cys3-Cys13, wherein the C-terminus of SEQ ID NO:29 is amidated;

[0466] {D-Tyr}-ICV-{Trp(Me)}-QDW-(Sar)-AHRCI-(AEEA)-GRKKRRQRRRPQ (SEQ ID NO: 30);

[0467] Where D-Tyr represents D-tyrosine, Trp(Me) represents N-methyltryptophan, Sar represents sarcosine, and AEEA represents 8-amino-3,6-dioxaoctanoyl (-NH-CH 2 CH 2 OCH 2 CH 2 -C(=O)-), and a disulfide bond is formed between Cys3-Cys13, wherein the C-terminus of SEQ ID NO: 30 is amidated;

[0468] {D-Tyr}-ICV-{Trp(Me)}-QDW-(Sar)-AHRCIRQIKIWFQNRR MKWKK (SEQ ID NO: 31);

[0469] wherein D-Tyr represents D-tyrosine, Trp(Me) represents N-methyltryptophan, Sar represents sarcosine, and a disulfide bond is formed between Cys3-Cys13, wherein the C-terminus of SEQ ID NO:31 is amidated;

[0470] {D-Tyr}-ICV-{Trp(Me)}-QDW-(Sar)-AHRCI-(AEEA)-RQIKIWFQNRRMKWKK(SEQ IDNO:32);

[0471] Where D-Tyr represents D-tyrosine, Trp(Me) represents N-methyltryptophan, Sar represents sarcosine, and AEEA represents 8-amino-3,6-dioxaoctanoyl (-NH-CH 2 CH 2 OCH 2 CH 2 -C(=O)-), and a disulfide bond is formed between Cys3-Cys13, wherein the C-terminus of SEQ ID NO:32 is amidated;

[0472] {D-Tyr}-ICV-{Trp(Me)}-QDW-(Sar)-AHRCI-(AEEA)-GRKKRRQRRRPQ(SEQ ID NO:33)

[0473] Where D-Tyr represents D-tyrosine, Trp(Me) represents N-methyltryptophan, Sar represents sarcosine, and AEEA represents 8-amino-3,6-dioxaoctanoyl (-NH-CH 2 CH 2 OCH 2 CH 2 -C(=O)-), and a disulfide bond is formed between Cys3-Cys13;

[0474] {D-Tyr}-ICV-{Trp(Me)}-QDW-(Sar)-AHRCI-(AEEA)-YGRKKRRQRRR(SEQ ID NO:34)

[0475] Where D-Tyr represents D-tyrosine, Trp(Me) represents N-methyltryptophan, Sar represents sarcosine, and AEEA represents 8-amino-3,6-dioxaoctanoyl (-NH-CH 2 CH 2 OCH 2 CH 2 -C(=O)-), and a disulfide bond is formed between Cys3-Cys13;

[0476] {D-Tyr}-ICV-{Trp(Me)}-QDW-(Sar)-AHRCI-(AEEA)-GYGRKKRRQRRR(SEQ ID NO:35)

[0477] Where D-Tyr represents D-tyrosine, Trp(Me) represents N-methyltryptophan, Sar represents sarcosine, and AEEA represents 8-amino-3,6-dioxaoctanoyl (-NH-CH 2 CH 2 OCH 2 CH 2 -C(=O)-), and a disulfide bond is formed between Cys3-Cys13; wherein the C-terminus of SEQ ID NO:35 is amidated;

[0478] {D-Tyr}-ICV-{Trp(Me)}-QDW-(Sar)-AHRCI-(AEEA)-GGYGRKKRRQRRR(SEQ ID NO:36)

[0479] Where D-Tyr represents D-tyrosine, Trp(Me) represents N-methyltryptophan, Sar represents sarcosine, and AEEA represents 8-amino-3,6-dioxaoctanoyl (-NH-CH 2 CH 2 OCH 2 CH 2 -C(=O)-), and a disulfide bond is formed between Cys3-Cys13; wherein the C-terminus of SEQ ID NO:36 is amidated;

[0480] {D-Tyr}-ICV-{Trp(Me)}-QDW-(Sar)-AHRCI-(AEEA)-GGGYGRKKRRQRRR(SEQ IDNO:37)

[0481] Where D-Tyr represents D-tyrosine, Trp(Me) represents N-methyltryptophan, Sar represents sarcosine, and AEEA represents 8-amino-3,6-dioxaoctanoyl (-NH-CH 2 CH 2 OCH 2 CH 2 -C(=O)-), and a disulfide bond is formed between Cys3-Cys13; wherein the C-terminus of SEQ ID NO:37 is amidated;

[0482] {ACI}-CV-{Trp(Me)}-QDWGAHRCT (SEQ ID NO: 38);

[0483] Wherein ACI represents acetylated isoleucine, Trp(Me) represents N-methyltryptophan, and a disulfide bond is formed between Cys2-Cys12;

[0484] {ACI}-CV-{Trp(Me)}-QDWGAHRCTGRKKRRQRRRPQ(SEQ ID NO:39)

[0485] wherein ACI represents acetylated isoleucine, Trp(Me) represents N-methyltryptophan, and a disulfide bond is formed between Cys2-Cys12, wherein the C-terminus of SEQ ID NO:39 is amidated;

[0486] {ACI}-CV-{Trp(Me)}-QDWGAHRCT-(AEEA)-GRKKRRQRRRPQ(SEQ ID NO:40)

[0487] ACI represents acetylated isoleucine, Trp(Me) represents N-methyltryptophan, and a disulfide bond is formed between Cys2 and Cys12. AEEA represents 8-amino-3,6-dioxaoctanoyl (-NH-CH 2 CH 2 OCH 2 CH 2 -C(=O)-), wherein the C-terminus of SEQ ID NO:40 is amidated;

[0488] {ACI}-CV-{Trp(Me)}-QDWGAHRCTRQIKIWFQNRRMKWKK(SEQ ID NO:41)

[0489] wherein ACI represents acetylated isoleucine, Trp(Me) represents N-methyltryptophan, and a disulfide bond is formed between Cys2-Cys12, wherein the C-terminus of SEQ ID NO:41 is amidated;

[0490] {ACI}-CV-{Trp(Me)}-QDWGAHRCT-(AEEA)-RQIKIWFQNRRMKWKK(SEQ ID NO:42)

[0491] ACI represents acetylated isoleucine, Trp(Me) represents N-methyltryptophan, and a disulfide bond is formed between Cys2 and Cys12. AEEA represents 8-amino-3,6-dioxaoctanoyl (-NH-CH 2 CH 2 OCH 2 CH 2 -C(=O)-), wherein the C-terminus of SEQ ID NO:42 is amidated;

[0492] {ACI}-CV-{Trp(Me)}-QDWGAHRCT-(AEEA)-GRKKRRQRRRPQ(SEQ ID NO:43)

[0493] ACI represents acetylated isoleucine, Trp(Me) represents N-methyltryptophan, and a disulfide bond is formed between Cys2 and Cys12. AEEA represents 8-amino-3,6-dioxaoctanoyl (-NH-CH 2 CH 2 OCH 2 CH 2 -C(=O)-);

[0494] {ACI}-CV-{Trp(Me)}-QDWGAHRCT-(AEEA)-YGRKKRRQRRR(SEQ ID NO:44)

[0495] ACI represents acetylated isoleucine, Trp(Me) represents N-methyltryptophan, and a disulfide bond is formed between Cys2 and Cys12. AEEA represents 8-amino-3,6-dioxaoctanoyl (-NH-CH 2 CH 2 OCH 2 CH 2 -C(=O)-), wherein the C-terminus of SEQ ID NO:44 is amidated;

[0496] {ACI}-CV-{Trp(Me)}-QDWGAHRCT-(AEEA)-GYGRKKRRQRRR(SEQ ID NO:45)

[0497] ACI represents acetylated isoleucine, Trp(Me) represents N-methyltryptophan, and a disulfide bond is formed between Cys2 and Cys12. AEEA represents 8-amino-3,6-dioxaoctanoyl (-NH-CH 2 CH 2 OCH 2 CH 2 -C(=O)-), wherein the C-terminus of SEQ ID NO:45 is amidated;

[0498] {ACI}-CV-{Trp(Me)}-QDWGAHRCT-(AEEA)-GGYGRKKRRQRRR(SEQ ID NO:46)

[0499] ACI represents acetylated isoleucine, Trp(Me) represents N-methyltryptophan, and a disulfide bond is formed between Cys2 and Cys12. AEEA represents 8-amino-3,6-dioxaoctanoyl (-NH-CH 2 CH 2 OCH 2 CH 2 -C(=O)-), wherein the C-terminus of SEQ ID NO:46 is amidated;

[0500] {ACI}-CV-{Trp(Me)}-QDWGAHRCT-(AEEA)-GGGYGRKKRRQRRR(SEQ ID NO:47)

[0501] ACI represents acetylated isoleucine, Trp(Me) represents N-methyltryptophan, and a disulfide bond is formed between Cys2 and Cys12. AEEA represents 8-amino-3,6-dioxaoctanoyl (-NH-CH 2 CH 2 OCH 2 CH 2 -C(=O)-), wherein the C-terminus of SEQ ID NO: 47 is amidated.

[0502] Test Example 1: C3 complement inhibition affinity test

[0503] The SCK method was used to determine the binding affinity of the peptide to C3 and C3b proteins. Peptide samples with increasing concentrations were injected into the human C3 or C3b protein channel immobilized on the biosensor chip. The obtained sensorgram was analyzed to determine the dissociation constant (KD), and the results showed the affinity of the peptide sample to bind to C3 and C3b.

[0504] The binding affinity and kinetic characteristics of Cp40 analogs were evaluated by SPR using a Biacore 3000 instrument (GE Healthcare, Piscataway Township, NJ). 18, 19, 47, 48 All experiments were performed at 25°C with 0.01 M HEPES, pH 7.4, 0.15 M NaCl, 3 mM EDTA, 0.005% surfactant P20 (HBSEP) as running buffer. Purified human C3b (Complement Technology, Inc., Tyler, TX) was coated on a CM5 sensor chip (GE Healthcare, Uppsala, Sweden) at a density of 11 000-20 000 resonance units (RUs) by amide coupling. An uncoated flow cell was used as a reference surface. The concentrations of each Cp40 mimetic were 2.5, 5, 10, 20, and 40 nM, respectively. Five samples were injected continuously at a flow rate of 30 μL / min, each sample was injected for 2 min, and the final dissociation step was 80 min. Cp40 was used as an internal reference for each run. Each peptide was screened in three independent experiments. The sensorgrams were processed using Scrubber software (BioLogic software, Campbell, Australia). The resulting data were fitted to a 1:1 Langmuir binding model in BIAevaluation software (GE Healthcare), and the equilibrium dissociation constant (KD) was obtained by the equation KD = KD / ka.

[0505] Table 1. The results of the peptide compounds of the present invention inhibiting affinity against C3 complement

[0506]

[0507]

[0508] The above results indicate that the polypeptide compound of the present invention has a strong inhibitory effect on C3 complement inhibition affinity.

[0509] Test Example 2: C3 complement inhibition test.

[0510] 1. The ability of Cp40 analogs to inhibit classical complement activation was evaluated by establishing an enzyme-linked immunosorbent assay (ELISA). 50 μL of 1% ovalbumin in PBS was applied to a 96-well plate at room temperature for 2 h.

[0511] 2. Seal the wells with 200 μL 1% BSA in PBS for 1 h, then coat with 50 μL 1:1000 diluted α-ovalbumin polyclonal antibody in PBS for 1 h. Wash three times with 200 μL 0.05% Tween 20 in PBS (PBS-t) at each step.

[0512] 3. Add serially diluted Cp40 analogs to a 96-well plate (VBS: 5 mM veronal, pH 7.4, containing 150 mM NaCl, 0.5 mM CaCl 2 and 0.5 mM MgCl 2 ), then human plasma (VBS 1:40) was added and incubated at room temperature for 15 min.

[0513] 4. After washing, use enzyme-labeled goat α-human C3 antibody (1:1000).

[0514] 5. Use HRP substrate (0.05% ABTS and 0.1% 30% H 2 O 2 Reactions were performed in 0.1 M sodium citrate, pH 4.2) and read at 405 nm using a VICTOR multilabel plate reader (PerkinElmer, Waltham, MA).

[0515] 6. Considering that 100% complement activation was obtained in the absence of C3 peptide inhibitor, the absorbance data obtained at 405 nm were converted to percentage of complement inhibition.

[0516] Test Example 3: Penetration of isolated rabbit cornea

[0517] The penetration of different liposomes through the isolated rabbit cornea was determined using a diffusion cell device. Rabbits were killed by intravenous injection of a lethal dose of sodium pentobarbital, the eyeballs were removed, and the cornea (together with the sclera of about 2 mm width around) was carefully taken out from the eyeballs and gently rinsed with PBS. The cornea was placed horizontally between the donor chamber and the receptor chamber, with the corneal epithelial surface facing the donor solution and fixed with a clamp. The donor solution was the sample solution, and the receptor solution was the PBS solution, and the bubbles were exhausted to make the corneal tissue fully contact with the receptor solution. During 4 hours, an equal amount of 100 μL sample was drawn from the receptor chamber every 0.5 h, and then an equal amount of PBS was immediately supplemented. The concentration of the extracted sample was measured using an ELISA test kit, and the apparent permeability coefficient was calculated.

[0518] The penetration of different liposomes through the isolated rabbit cornea was determined using a diffusion cell device. Rabbits were killed by intravenous injection of a lethal dose of sodium pentobarbital, the eyeballs were removed, and the cornea (together with the sclera of about 2 mm width around) was carefully taken out from the eyeballs and gently rinsed with PBS. The cornea was placed horizontally between the donor chamber and the receptor chamber, with the corneal epithelial surface facing the donor solution and fixed with a clamp. The donor solution was the sample solution, and the receptor solution was the PBS solution, and the bubbles were exhausted to make the corneal tissue fully contact with the receptor solution. During 4 hours, an equal amount of 100 μL sample was drawn from the receptor chamber every 0.5 h, and then an equal amount of PBS was immediately supplemented. The concentration of the extracted sample was measured using an ELISA test kit, and the apparent permeability coefficient was calculated.

[0519] Table 2. Penetration of the polypeptide compound of SEQ ID NO: 29 of the present invention against the isolated rabbit cornea

[0520]

[0521] The polypeptide compound of the present invention has certain penetrability to the isolated rabbit cornea.

[0522] Test Example 4: Study on the intraocular distribution of peptides

[0523] (1) Experimental groups: PBS blank control group, control C3 complement inhibitor peptide group, Penetratin peptide group, and transmembrane C3 complement inhibitor group.

[0524] (2) Administration method: After intraperitoneal injection of sodium pentobarbital, SD rats were anesthetized. For the eye drop group, 10 μL of FITC-labeled peptide solution or PBS was sucked up with a pipette tip, and the eyelids were gently closed to allow the drug to be evenly distributed. The drug was administered once every 10 minutes for a total of 6 times.

[0525] (3) Preparation of frozen sections: After the above administration, rats were killed at different time points of 0.5h, 3h, 6h, and 24h, and the eyeballs were removed and rinsed with PBS three times. After that, they were fixed in 4% paraformaldehyde at 4°C overnight, and then dehydrated in 10%, 20%, and 30% sucrose solutions at 4°C, with each concentration dehydrating for 12h. After fixation and dehydration, OCT was embedded, and the specimens were placed in a -20°C cryostat, and l0μm sections were made to prepare frozen sections of eye tissue.

[0526] (4) Observation of the intraocular distribution of peptides and semi-quantitative determination of FITC fluorescence intensity: A confocal laser scanning microscope or fluorescence microscope equipped with a fluorescence filter system was used to observe the intraocular distribution of FITC-labeled peptides and take photos. The fluorescence intensity of each group of frozen sections was evaluated according to four levels based on the signal strength: level 0, only background fluorescence; level 1, weak fluorescence; level 2, moderate fluorescence; level 3, strong fluorescence.

Claims

1. A polypeptide comprising or consisting of the amino acid sequence of SEQ ID NO: 1 or SEQ ID NO: 11 as shown below: X1ICVX2QDWX3AHRCI (SEQ ID NO: 1), wherein X1 represents D-tyrosine; X2 is tryptophan or a tryptophan analogue, wherein the tryptophan analogue is selected from: a) N-methyltryptophan; b) tryptophan having a C1-C4 alkyl substituent at the 5-position of the indole ring; c) 5-fluorotryptophan; or d) 6-fluorotryptophan; X3 represents sarcosine; wherein a disulfide bond is formed between the cysteine ​​residues at positions 3 and 13; or X4CVX2QDWGAHRCT (SEQ ID NO: 11), wherein X2 is tryptophan or a tryptophan analogue selected from a) N-methyltryptophan; b) tryptophan having a C1-C4 alkyl substituent at the 5-position of the indole ring; c) 5-fluorotryptophan; or d) 6-fluorotryptophan; X4 indicates acetylated isoleucine; A disulfide bond is formed between the 2nd and 12th cysteines.

2. A fusion polypeptide comprising: A) a polypeptide represented by SEQ ID NO: 1, X1ICVX2QDWX3AHRCI (SEQ ID NO: 1), wherein X1 represents D-tyrosine; X2 is selected from tryptophan or a tryptophan analog, wherein the tryptophan analog is selected from: a) N-methyltryptophan; b) tryptophan having a C1-C4 alkyl substituent at the 5-position of the indole ring; c) 5-fluorotryptophan; or d) 6-fluorotryptophan; X3 represents sarcosine; wherein a disulfide bond is formed between the cysteine ​​residues at positions 3 and 13; and B) A polypeptide comprising any of the following amino acid sequences: GRKKRRQRRRPQ (SEQ ID NO: 21); RQIKIWFQNRRMKWKK (SEQ ID NO: 22); GRKKRRQRRRPQ (SEQ ID NO: 23); YGRKKRRQRRR (SEQ ID NO: 24); GYGRKKRRQRRR (SEQ ID NO: 25); GGYGRKKRRQRRR (SEQ ID NO: 26); or GGGYGRKKRRQRRR (SEQ ID NO: 27); Optionally, the polypeptide shown in any one of SEQ ID NO: 21 to SEQ ID NO: 27 has or does not have C-terminal amidation; Preferably, the polypeptide shown in any one of SEQ ID NO: 21 to SEQ ID NO: 27 has a membrane-penetrating function; Preferably, the polypeptide shown in any one of SEQ ID NO: 21 to SEQ ID NO: 27 is located at the C-terminus of the polypeptide shown in SEQ ID NO:

1.

3. The fusion polypeptide according to claim 2, wherein the polypeptide represented by SEQ ID NO: 1 is directly linked to the polypeptide represented by any one of SEQ ID NO: 21 to SEQ ID NO: 27, or is linked via a linker.

4. The fusion polypeptide according to claim 2, wherein the polypeptide represented by SEQ ID NO: 1 is connected to the polypeptide represented by any one of SEQ ID NO: 21 to SEQ ID NO: 27 via a linker, and preferably, the linker is a fragment represented by AEEA; Wherein AEEA represents 8-amino-3,6-dioxaoctanoyl (-NH-CH2CH2OCH2CH2-C(=O)-).

5. A fusion polypeptide comprising: A) a polypeptide represented by SEQ ID NO: 11, X4CVX2QDWGAHRCT (SEQ ID NO: 11), wherein X2 is tryptophan or a tryptophan analogue selected from a) N-methyltryptophan; b) tryptophan having a C1-C4 alkyl substituent at the 5-position of the indole ring; c) 5-fluorotryptophan; or d) 6-fluorotryptophan; X4 indicates acetylated isoleucine; wherein a disulfide bond is formed between the cysteine ​​residues at positions 2 and 12; and B) a polypeptide comprising the following amino acid sequence: GRKKRRQRRRPQ (SEQ ID NO: 21); RQIKIWFQNRRMKWKK (SEQ ID NO: 22); GRKKRRQRRRPQ (SEQ ID NO: 23); YGRKKRRQRRR (SEQ ID NO: 24); GYGRKKRRQRRR (SEQ ID NO: 25); GGYGRKKRRQRRR (SEQ ID NO: 26); or GGGYGRKKRRQRRR (SEQ ID NO: 27); Optionally, the polypeptide shown in any one of SEQ ID NO: 21 to SEQ ID NO: 27 has or does not have C-terminal amidation; Preferably, the polypeptide shown in any one of SEQ ID NO: 21 to SEQ ID NO: 27 has a membrane-penetrating function; Preferably, the polypeptide shown in any one of SEQ ID NO:21 to SEQ ID NO:27 is located at the C-terminus of the polypeptide shown in SEQ ID NO:

11.

6. The fusion polypeptide according to claim 5, wherein the polypeptide represented by SEQ ID NO: 11 is directly linked to the polypeptide represented by any one of SEQ ID NO: 21 to SEQ ID NO: 27, or is linked via a linker.

7. The fusion polypeptide according to claim 5, wherein the polypeptide represented by SEQ ID NO: 11 is connected to the polypeptide represented by any one of SEQ ID NO: 21 to SEQ ID NO: 27 via a linker, preferably, the linker is a fragment represented by AEEA; Here, AEEA represents 8-amino-3,6-dioxaoctanoyl (-NH-CH2CH2OCH2CH2-C(=O)-).

8. The polypeptide according to claim 1, or the fusion polypeptide according to any one of claims 2 to 7, which has an acetylation modification at the N-terminus and / or an amidation modification at the C-terminus.

9. The polypeptide according to claim 1 or 8, or the fusion polypeptide according to any one of claims 2 to 8, which has an amino acid sequence as shown in any one of SEQ ID NOs: 2 to 10 and 12 to 20, or has at least 75% sequence identity thereto; or has at most 3, preferably at most 2 or at most 1 amino acid sequence substitutions, deletions or additions thereto, preferably, The polypeptide according to claim 1 or 8, or the fusion polypeptide according to any one of claims 2 to 8, having an amino acid sequence as shown in any one of SEQ ID NOs: 28 to 47: {D-Tyr}-ICV-{Trp(Me)}-QDW-(Sar)-AHRCI, (SEQ ID NO: 28) wherein D-Tyr represents D-tyrosine, Trp(Me) represents N-methyltryptophan, Sar represents sarcosine, and a disulfide bond is formed between Cys3-Cys13; {D-Tyr}-ICV-{Trp(Me)}-QDW-(Sar)-AHRCIGRKKRRQRRRPQ(SEQ ID NO: 29) wherein D-Tyr represents D-tyrosine, Trp(Me) represents N-methyltryptophan, Sar represents sarcosine, and a disulfide bond is formed between Cys3-Cys13, wherein the C-terminus of SEQ ID NO:29 is amidated; {D-Tyr}-ICV-{Trp(Me)}-QDW-(Sar)-AHRCI-(AEEA)-GRKKRRQRRRPQ(SEQ ID NO: 30) wherein D-Tyr represents D-tyrosine, Trp(Me) represents N-methyltryptophan, Sar represents sarcosine, AEEA represents 8-amino-3,6-dioxaoctanoyl (-NH-CH2CH2OCH2CH2-C(=O)-), and a disulfide bond is formed between Cys3-Cys13, wherein the C-terminus of SEQ ID NO:30 is amidated; {D-Tyr}-ICV-{Trp(Me)}-QDW-(Sar)-AHRCIRQIKIWFQNRR MKWKK (SEQ ID NO: 31) wherein D-Tyr represents D-tyrosine, Trp(Me) represents N-methyltryptophan, Sar represents sarcosine, and a disulfide bond is formed between Cys3-Cys13, wherein the C-terminus of SEQ ID NO:31 is amidated; {D-Tyr}-ICV-{Trp(Me)}-QDW-(Sar)-AHRCI-(AEEA)-RQIKIWFQNRRMKWKK(SEQ ID NO:32) wherein D-Tyr represents D-tyrosine, Trp(Me) represents N-methyltryptophan, Sar represents sarcosine, AEEA represents 8-amino-3,6-dioxaoctanoyl (-NH-CH2CH2OCH2CH2-C(=O)-), and a disulfide bond is formed between Cys3-Cys13, wherein the C-terminus of SEQ ID NO:32 is amidated; {D-Tyr}-ICV-{Trp(Me)}-QDW-(Sar)-AHRCI-(AEEA)-GRKKRRQRRRPQ(SEQ ID NO:33) wherein D-Tyr represents D-tyrosine, Trp(Me) represents N-methyltryptophan, Sar represents sarcosine, AEEA represents 8-amino-3,6-dioxaoctanoyl (-NH-CH2CH2OCH2CH2-C(=O)-), and a disulfide bond is formed between Cys3-Cys13; {D-Tyr}-ICV-{Trp(Me)}-QDW-(Sar)-AHRCI-(AEEA)-YGRKKRRQRRR(SEQ ID NO:34) wherein D-Tyr represents D-tyrosine, Trp(Me) represents N-methyltryptophan, Sar represents sarcosine, AEEA represents 8-amino-3,6-dioxaoctanoyl (-NH-CH2CH2OCH2CH2-C(=O)-), and a disulfide bond is formed between Cys3-Cys13; {D-Tyr}-ICV-{Trp(Me)}-QDW-(Sar)-AHRCI-(AEEA)-GYGRKKRRQRRR(SEQ ID NO:35) wherein D-Tyr represents D-tyrosine, Trp(Me) represents N-methyltryptophan, Sar represents sarcosine, AEEA represents 8-amino-3,6-dioxaoctanoyl (-NH-CH2CH2OCH2CH2-C(=O)-), and a disulfide bond is formed between Cys3-Cys13; wherein the C-terminus of SEQ ID NO:35 is amidated; {D-Tyr}-ICV-{Trp(Me)}-QDW-(Sar)-AHRCI-(AEEA)-GGYGRKKRRQRRR(SEQ ID NO:36) wherein D-Tyr represents D-tyrosine, Trp(Me) represents N-methyltryptophan, Sar represents sarcosine, AEEA represents 8-amino-3,6-dioxaoctanoyl (-NH-CH2CH2OCH2CH2-C(=O)-), and a disulfide bond is formed between Cys3-Cys13; wherein the C-terminus of SEQ ID NO:36 is amidated; {D-Tyr}-ICV-{Trp(Me)}-QDW-(Sar)-AHRCI-(AEEA)-GGGYGRKKRRQRRR(SEQ ID NO:37) wherein D-Tyr represents D-tyrosine, Trp(Me) represents N-methyltryptophan, Sar represents sarcosine, AEEA represents 8-amino-3,6-dioxaoctanoyl (-NH-CH2CH2OCH2CH2-C(=O)-), and a disulfide bond is formed between Cys3-Cys13; wherein the C-terminus of SEQ ID NO:37 is amidated; {ACI}-CV-{Trp(Me)}-QDWGAHRCT(SEQ ID NO: 38) Wherein ACI represents acetylated isoleucine, Trp(Me) represents N-methyltryptophan, and a disulfide bond is formed between Cys2-Cys12; {ACI}-CV-{Trp(Me)}-QDWGAHRCT GRKKRRQRRRPQ(SEQ ID NO:39) wherein ACI represents acetylated isoleucine, Trp(Me) represents N-methyltryptophan, and a disulfide bond is formed between Cys2-Cys12, wherein the C-terminus of SEQ ID NO:39 is amidated; {ACI}-CV-{Trp(Me)}-QDWGAHRCT-(AEEA)-GRKKRRQRRRPQ(SEQ ID NO:40) wherein ACI represents acetylated isoleucine, Trp(Me) represents N-methyltryptophan, and a disulfide bond is formed between Cys2-Cys12, and AEEA represents 8-amino-3,6-dioxaoctanoyl (-NH-CH2CH2OCH2CH2-C(=O)-), wherein the C-terminus of SEQ ID NO:40 is amidated; {ACI}-CV-{Trp(Me)}-QDWGAHRCTRQIKIWFQNRRMKWKK(SEQ ID NO:41) wherein ACI represents acetylated isoleucine, Trp(Me) represents N-methyltryptophan, and a disulfide bond is formed between Cys2-Cys12, wherein the C-terminus of SEQ ID NO:41 is amidated; {ACI}-CV-{Trp(Me)}-QDWGAHRCT-(AEEA)-RQIKIWFQNRRMKWKK(SEQ ID NO:42) wherein ACI represents acetylated isoleucine, Trp(Me) represents N-methyltryptophan, and a disulfide bond is formed between Cys2-Cys12, and AEEA represents 8-amino-3,6-dioxaoctanoyl (-NH-CH2CH2OCH2CH2-C(=O)-), wherein the C-terminus of SEQ ID NO:42 is amidated; {ACI}-CV-{Trp(Me)}-QDWGAHRCT-(AEEA)-GRKKRRQRRRPQ(SEQ ID NO:43) Wherein ACI represents acetylated isoleucine, Trp(Me) represents N-methyltryptophan, and a disulfide bond is formed between Cys2-Cys12, and AEEA represents 8-amino-3,6-dioxaoctanoyl (-NH-CH2CH2OCH2CH2-C(=O)-); {ACI}-CV-{Trp(Me)}-QDWGAHRCT-(AEEA)-YGRKKRRQRRR(SEQ ID NO:44) wherein ACI represents acetylated isoleucine, Trp(Me) represents N-methyltryptophan, and a disulfide bond is formed between Cys2-Cys12, and AEEA represents 8-amino-3,6-dioxaoctanoyl (-NH-CH2CH2OCH2CH2-C(=O)-), wherein the C-terminus of SEQ ID NO:44 is amidated; {ACI}-CV-{Trp(Me)}-QDWGAHRCT-(AEEA)-GYGRKKRRQRRR(SEQ ID NO:45) wherein ACI represents acetylated isoleucine, Trp(Me) represents N-methyltryptophan, and a disulfide bond is formed between Cys2-Cys12, and AEEA represents 8-amino-3,6-dioxaoctanoyl (-NH-CH2CH2OCH2CH2-C(=O)-), wherein the C-terminus of SEQ ID NO:45 is amidated; {ACI}-CV-{Trp(Me)}-QDWGAHRCT-(AEEA)-GGYGRKKRRQRRR(SEQ ID NO:46) wherein ACI represents acetylated isoleucine, Trp(Me) represents N-methyltryptophan, and a disulfide bond is formed between Cys2-Cys12, and AEEA represents 8-amino-3,6-dioxaoctanoyl (-NH-CH2CH2OCH2CH2-C(=O)-), wherein the C-terminus of SEQ ID NO:46 is amidated; {ACI}-CV-{Trp(Me)}-QDWGAHRCT-(AEEA)-GGGYGRKKRRQRRR(SEQ ID NO:47) Wherein ACI represents acetylated isoleucine, Trp(Me) represents N-methyltryptophan, and a disulfide bond is formed between Cys2-Cys12, and AEEA represents 8-amino-3,6-dioxaoctanoyl (-NH-CH2CH2OCH2CH2-C(=O)-), wherein the C-terminus of SEQ ID NO:47 is amidated.

10. The polypeptide according to any one of claims 1, 8 or 9, or the fusion polypeptide according to any one of claims 2 to 9, which is a C3 complement inhibitor, preferably, which is capable of penetrating cell membranes or body tissue barriers.

11. A pharmaceutical composition comprising the polypeptide according to any one of claims 1, 8 or 9, or the fusion polypeptide according to any one of claims 2-9, and one or more pharmaceutically acceptable carriers or excipients.

12. Use of the polypeptide according to any one of claims 1, 8 or 9, the fusion polypeptide according to any one of claims 2-9, or the pharmaceutical composition according to claim 11 for preparing a medicament for treating a disease, especially an eye disease, preferably, The ocular disease is selected from age-related macular degeneration (AMD), diabetic macular edema (DME), inflammatory eye diseases, optic neuropathy eye diseases and ocular microangiopathy-related disorders, preferably, the ocular microangiopathy-related disorders are selected from diabetic retinopathy (DR), diabetic macular edema (DME), retinal vein occlusion (RVO), central retinal vein occlusion (CRVO), macular degeneration, retinopathy of prematurity (ROP), neovascular glaucoma, retinitis pigmentosa (RP), proliferative and non-proliferative retinopathy, retinal angioma The present invention relates to a diabetic retinal neovascularization, wherein the retinal neovascularization is caused by iris neovascularization, intraocular neovascularization, corneal neovascularization, retinal neovascularization, polypoidal choroidal vasculopathy (PCV), choroidal neovascularization, retinal degeneration, retinal vascular permeability associated with diabetic retinopathy and diabetic macular edema; in particular, the present invention is selected from diabetic retinopathy (DR), diabetic macular edema (DME), retinal vein occlusion (RVO), central retinal vein occlusion (CRVO) and wet age-related macular degeneration (wet AMD).

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