Peptide inhibitors and uses thereof

Abstract

The present invention relates to a peptide inhibitor for inhibiting activation of TGF-beta1 in which TSP-1 participates or a pharmaceutically acceptable salt, solvate or prodrug thereof, a pharmaceutical composition comprising the peptide or the pharmaceutically acceptable salt, solvate or prodrug thereof, and a method for treating or preventing TGF-beta1 related diseases, in particular fibrosis and solid tumors, using the peptide inhibitor.

Description

Peptide inhibitors and uses thereof Technical Field

[0001] The present invention relates to peptide inhibitors that inhibit TSP-1-involved TGF-β1 activation or pharmaceutically acceptable salts, solvates or prodrugs thereof, as well as pharmaceutical compositions comprising the peptides or pharmaceutically acceptable salts, solvates or prodrugs thereof and methods for using the same to treat or prevent TGF-β1-related diseases, especially fibrosis and solid tumors. Background Art

[0002] Fibrosis is a pathological process characterized by the abnormal, excessive proliferation of fibrous connective tissue during injury and repair, resulting in scarring. It is characterized by excessive deposition of the extracellular matrix (ECM). Fibrosis not only leads to irreversible decline in tissue and organ function but, in severe cases, can impact patients' quality of life and even be life-threatening. Furthermore, fibrosis promotes tumorigenesis and distant metastasis, fosters the formation of an immunosuppressive tumor microenvironment, and can lead to immunotherapy resistance or immune nonresponse.

[0003] Transforming growth factor β (TGFβ) is a key regulatory molecule in the development and progression of fibrosis and tumors. TGF-β and its signaling pathway have been proposed as therapeutic targets for fibrotic disorders and tumors. However, as a multifunctional cytokine, TGF-β is also involved in a variety of normal physiological processes, resulting in high toxicity of drugs that directly target TGF-β and its signaling pathway. Currently, some such drug development projects have been slowed or even terminated after entering the clinical stage due to severe drug-related adverse reactions.

[0004] During synthesis and secretion, TGF-β begins as an inactive precursor structure. It must undergo a series of activation events to become an active fragment before it can bind to its receptor and mediate downstream pathway transduction. TGF-β activation occurs under different environmental and environmental conditions and involves different protein molecules. Therefore, targeting specific proteins involved in disease activation has become a new research direction to overcome the difficulties in developing TGF-β drugs.

[0005] Thrombospondin-1 (TSP-1) is a protein that has been clinically found to be actively expressed in fibrosis and tumor-related indications. Studies have shown that TSP-1 can participate in TGF-β activation events in chronic inflammation and fibrosis and is the main activator of TGF-β1 in vivo (Cell, Vol. 93, 1159–1170, June 26, 1998, Copyright 1998 by Cell Press). Considering clinical applications, peptide drugs targeting TGF-β activation may offer a safer strategy than other therapeutic anti-TGF-β1 molecules due to their ability to act at the site of TGF-β1 activation, ease of catabolism, and low antigenicity. Therefore, the present invention proposes to design peptide components targeting the TSP-1 domain involved in TGF-β activation, in the hope of providing new peptide drugs to improve fibrosis and cancer.

[0006] SUMMARY OF THE INVENTION

[0007] Through meticulous research, the present inventors unexpectedly discovered a group of short peptides of 5-13 amino acids containing an amino acid motif (DXED) that not only effectively inhibit the interaction between the TSP-1 receptor and the TSP-1 / L-TGF-β1 complex, but also effectively block TSP-1-dependent L-TGF-β1 activation and subsequent TGF-β signaling. Based on this discovery, the present inventors developed the inhibitory peptides of the present invention (hereinafter referred to as "TGF-β1 activation inhibitory peptides"). As shown in the Examples, the inhibitory peptides of the present invention exhibit TGF-β1 activation inhibitory activity in both cyclic and linear forms. Furthermore, compared to a similar peptide known in the prior art (SEQ ID NO:50), the inhibitory peptides exhibit a more potent inhibitory effect on TGF-β1 activation while being shortened to achieve greater cost-effectiveness. In some preferred embodiments, the inhibitory peptides of the present invention also exhibit improved solubility, stability, and / or bioavailability compared to the known similar peptides.

[0008] Furthermore, the inhibitory peptides of the present invention also exhibit favorable drug safety advantages. Studies on TSP-1 have shown that it contains multiple functional domains that can independently bind to different cell surface receptors and participate in various physiological processes such as hemostasis, cell adhesion, cell migration, apoptosis, and anti-angiogenesis. Due to their small molecular weight, high activity, and design targeting the specific domain on TSP-1 involved in TGF-β1 activation, the inhibitory peptides of the present invention can effectively reduce interference with the remaining functional domains of TSP-1 and related physiological functions, ensuring the specificity and safety of the drug's action.

[0009] Therefore, in a first aspect, the present invention provides a TGF-β1 activation inhibitory peptide or a pharmaceutically acceptable salt, solvate or prodrug thereof. The TGF-β1 activation inhibitory peptide according to the present invention can specifically block TSP-1-involved TGF-β1 activation and has the following general formula (I) amino acid sequence: R 1 -X 1 -X 2 -X 3 -X 4 -X 5-X 6 -X 7 -R 2 (I),

[0010] in,

[0011] R 1 is acetyl or absent;

[0012] R 2 It is amino or does not exist;

[0013] X 1 Not present, or selected from T and S;

[0014] X 2 Selected from natural amino acids or non-natural amino acids, preferably, selected from polar side chain amino acids, such as uncharged polar side chain amino acids S, C, G, N, Q, T, Y or negatively charged polar side chain amino acids D and E, more preferably selected from amino acids N, C, Q, S, T, E and D; more preferably selected from amino acids Q, C, N, E or S;

[0015] X 3 It is D;

[0016] X 4 is selected from amino acids with small side chains, such as A, C, G, P, S, T and V, more preferably selected from amino acids A, P and S, and more preferably selected from amino acids A and P;

[0017] X 5 It is E;

[0018] X 6 It is D;

[0019] X 7 does not exist, or is selected from the amino acid sequence Z 1 Z 2 Z 3 SZ 4 LQ is a sequence of 1, 2, 3, 4, 5, 6 or 7 amino acids from the amino terminus to the carboxyl terminus.

[0020] where Z 1 is the amino acid N, Q, P, A, L, V, M, or I, preferably N, P, or L, most preferably N or P;

[0021] Z 2 is an amino acid T, S, V, C, A, K, or R, preferably T, C or K, more preferably, when Z 2 When it is amino acid C or K, Z 2 With X 2 or X 3 The amino acid side chains form covalent bonds,

[0022] Z 3 is amino acid A, V, L, or I, preferably A or V,

[0023] Z 4 is amino acid F, Y, H, or W, preferably F or H;

[0024] Optionally, when X 4 When A, at X 3 and X 4 0-1 small side chain amino acids, such as P or G, are inserted between, preferably, P is inserted;

[0025] Optionally, when X 7 When the peptide is composed of 1-5 amino acids, the peptide is 7 The residues may be followed by 0-2 additional amino acid residues, for example, 1-2 amino acid residues selected from the group consisting of Q, E, D, and N, or 1 amino acid residue selected from the group consisting of C or K.

[0026] In some embodiments, in the peptides of the present invention,

[0027] X 1 Not present, or selected from T and S;

[0028] X 2 selected from the group consisting of amino acids Q, N, C, S, T, E and D;

[0029] X 3 It is D;

[0030] X 4 Selected from A, C, G, P, S, T and V;

[0031] X 5 It is E;

[0032] X 6 It is D;

[0033] X 7 does not exist, or is selected from the amino acid sequence Z 1 Z 2 Z 3 SZ 4 LQ is a sequence of 1, 2, 3, 4, 5, 6 or 7 amino acids from the amino terminus to the carboxyl terminus, wherein

[0034] Z 1 is N, P or L;

[0035] Z 2 It is T or C or K;

[0036] Z 3 is A or V;

[0037] Z 4 It is F, Y, H, or W.

[0038] In some embodiments, in the peptides of the present invention,

[0039] X 1 Selected from T and S;

[0040] X 2 Selected from Q and N;

[0041] X 3 It is D;

[0042] X 4 Selected from A and P;

[0043] X 5 It is E;

[0044] X 6 It’s D.

[0045] In some embodiments, X 7 Does not exist.

[0046] In other embodiments, X 7 By Z 1 , Z 1 Z 2 , Z 1 Z 2 Z 3 , Z 1 Z 2 Z 3 S, Z 1 Z 2 Z 3 SZ 4 , Z 1 Z 2 Z 3 SZ 4 L or Z 1 Z 2 Z 3 SZ 4 LQ composition, preferably, X 7 By Z 1 Z 2 Z 3 , Z 1 Z 2 Z 3 S or Z 1 Z 2 Z 3 SZ 4 composition,

[0047] Among them, preferably

[0048] Z1 N, P or L, preferably N or P;

[0049] Z 2 is T, K or C;

[0050] Z 3 A or V, preferably A;

[0051] Z 4 is F, Y, H, or W, preferably F or H.

[0052] The peptides of the present invention may be linear peptides or cyclized peptides.

[0053] In a second aspect, the present invention also provides the therapeutic and / or preventive use of the inhibitory peptide of the present invention or a pharmaceutically acceptable salt, solvate or prodrug thereof in diseases caused by TSP-1-involved TGF-β1 activation or diseases related thereto (i.e., "TGF-β1-related diseases"). The present invention also provides the use of the inhibitory peptide of the present invention or a pharmaceutically acceptable salt, solvate or prodrug thereof in the preparation of a drug for preventing or treating fibrotic disorders. The fibrotic disorder is characterized by TGF-β1 activation involving TSP-1, and exhibits excessive deposition of the extracellular matrix (ECM), but is not limited to the site of disease occurrence, nor is it limited to traditional disease classifications. The present invention also provides the use of the inhibitory peptide of the present invention in the preparation of a drug for preventing or treating cancers associated with TGF-β1 activation.

[0054] BRIEF DESCRIPTION OF THE DRAWINGS

[0055] Figure 1 shows HE-stained pathological sections of a mouse model of fibrosis. HE staining was used to assess the severity of inflammation, demonstrating the inhibitory effect of the polypeptide of the present invention on inflammatory infiltration in pathological sites of fibrosis.

[0056] Figure 2 shows the MASSON'S stained pathological sections of a fibrosis mouse disease model. The severity of fibrosis was assessed by MASSON'S trichrome staining, demonstrating the inhibitory effect of the polypeptide of the present invention on fibrosis deposition in pathological sites of fibrosis.

[0057] FIG3 shows the MASSON'S stained pathological sections of a fibrosis rat disease model, demonstrating the inhibitory effect of the polypeptide of the present invention on fibrosis deposition in pathological sites of fibrosis.

[0058] Summary of the Invention

[0059] definition

[0060] In order to make the present invention more easily understood, certain scientific and technical terms are specifically defined below. Unless otherwise specifically defined elsewhere herein, the scientific and technical terms used herein have the meanings commonly understood by those of ordinary skill in the art to which the present invention belongs. The abbreviations for amino acid residues are standard three-letter and / or one-letter codes used in the art to refer to one of the 20 commonly used amino acids. Unless otherwise indicated, amino acid sequences are written from left to right in the direction from amino group to carboxyl group.

[0061] As used herein (including in the claims), singular forms include their corresponding plural forms unless the context clearly dictates otherwise.

[0062] The term "about" refers to a value that is within an acceptable error range for a particular value as determined by one of ordinary skill in the art, where the error range depends on the measurement means used to measure or determine the value, i.e., the limitations of the measurement system. For example, "about" can mean within 1 or more standard deviations, as practiced in the art. Alternatively, "about" can mean a range of up to 5%, 10%, or 20% (i.e., ±5%, ±10%, or ±20%).

[0063] The term "and / or" when used to link two or more alternatives should be understood to mean any one of the alternatives or any two or more of the alternatives.

[0064] As used herein, the term "comprising" or "including" means including the recited elements, integers, or steps, but not excluding any other elements, integers, or steps. In this document, when the term "comprising" or "including" is used, unless otherwise indicated, it also encompasses the situation of consisting of the recited elements, integers, or steps. For example, when it is mentioned that a particular sequence is "comprising", it is also intended to encompass a peptide or polypeptide consisting of the particular sequence.

[0065] As used herein, the terms "peptide" and "polypeptide" are used interchangeably and refer to an amino acid sequence between 2 and 100 amino acids in length, wherein the amino acids are linked by peptide bonds. Amino acids can be naturally occurring or non-naturally occurring.

[0066] As used herein, the term "conservative amino acid substitution" or "conservative amino acid substitution" means an amino acid substitution that does not adversely affect or alter the biological function of the polypeptide comprising the amino acid sequence. A typical conservative amino acid substitution refers to the substitution of one amino acid with another amino acid having similar chemical properties (e.g., charge or hydrophobicity). Conservative substitution tables for functionally similar amino acids are well known in the art. In the present invention, conservative substitution residues can be from the following conservative substitution table, particularly the preferred conservative amino acid substitution residues in the table.

[0067] Detailed Description of the Invention

[0068] During their research, the inventors surprisingly discovered that a short peptide with a negatively charged amino acid motif (DXED) can effectively inhibit TSP-1-dependent TGF-β1 activation. Through structure-activity relationship and amino acid substitution analysis, the inventors further identified essential amino acids and potential substitution sites within the peptide sequence. Furthermore, through peptide cyclization and / or backbone modification, the inventors optimized the structure and developed a series of novel TGF-β1 activation inhibitory peptides with improved biological activity and / or physicochemical properties.

[0069] Inhibitory peptides of the present invention

[0070] In one aspect, therefore, the present invention provides a TGF-β1 activation inhibitory peptide or a pharmaceutically acceptable salt, solvate or prodrug thereof. The peptide of the present invention comprises a core motif rich in negative charge (DXED) and is 5-13 amino acids long. Preferably, the peptide of the present invention consists of 7-11 amino acids. In some embodiments, the peptide of the present invention is a linear or cyclized pentapeptide or hexapeptide, such as a linear or first-stage cyclized hexapeptide. In some embodiments, the peptide of the present invention is a linear or cyclized heptapeptide, such as a head-to-tail cyclized heptapeptide. In other embodiments, the peptide of the present invention is a linear or cyclized octapeptide, especially an octapeptide with a side chain cyclized, preferably a disulfide bond cyclized. In other embodiments, the peptide of the present invention is a linear or cyclized nonapeptide, especially a linear nonapeptide. In other embodiments, the peptide of the present invention is a linear or cyclized decapeptide, such as a head-to-tail cyclized decapeptide. In other embodiments, the peptide of the present invention is a linear or cyclized undecapeptide, a dodecapeptide or a dodecapeptide, preferably a linear undecapeptide.

[0071] In some embodiments, the peptides of the present invention have 0-5 amino acid differences, e.g., 0-2 amino acid differences, between the full length of the peptide of the present invention and the corresponding portion of SEQ ID NO:51 aligned thereto after maximum alignment. Such amino acid differences include amino acid substitutions, additions, and deletions. For example, the peptides of the present invention may be identical to the sequence of SEQ ID NO:51 over the corresponding portion, or may have 1 or 2 amino acid substitutions, or may have 1-2 amino acid additions at the N-terminus or C-terminus, particularly at the C-terminus. The substituting amino acids may be natural or unnatural amino acids. In some embodiments, the peptides of the present invention comprise a proline substitution, preferably at the second residue position of the core sequence DXED; or at the first residue position immediately adjacent to the C-terminus of the core sequence. In some embodiments, the overall net charge of the peptides of the present invention is negative, e.g., -1, -2, -3, or -4, preferably -2 or -3.

[0072] In some embodiments, the peptide comprises a core amino acid sequence selected from the group consisting of: DAED or DPED. In other embodiments, the peptide further comprises an N or P residue located at the C-terminus of the core sequence, such that the peptide comprises an amino acid sequence selected from the group consisting of: DAEDN; DPEDN; or DAEDP.

[0073] In some embodiments, the peptides of the present invention are cyclic peptides. In some embodiments, the peptides of the present invention can be cyclized by covalently linking the first and last amino acids. In other embodiments, the peptides of the present invention can be cyclized by forming a lactam bond or a disulfide bond between two amino acid side chains. To this end, suitable amino acids or analogs with appropriate side chain groups can be placed in appropriate positions of the peptides of the present invention to promote the formation of a lactam bond or a disulfide bond in the peptide. In some embodiments, the lactam is formed by coupling the side chain amino functional group of the amino acid residue at the C-terminus of the core motif (DXED) of the peptides of the present invention with the carboxylic acid group of the amino acid at the N-terminus of the core motif (DXED). In some embodiments, the disulfide bond is formed by oxidative coupling of a cysteine ​​at the C-terminus of the core motif (DXED) with a cysteine ​​at the N-terminus of the core motif (DXED). In some embodiments of the lactam cyclic peptide, the side chain carboxyl group of the first aspartic acid (D) of the core motif or the aspartic acid (D) or glutamic acid (E) located at its N-terminus forms a lactam bond with the side chain amino group of the lysine (K) located at the C-terminus of the core motif, and the two amino acids forming the ring are separated by no less than 4 amino acids, for example, 4, 5 or 6 amino acids. Preferably, the lysine residue forming the ring is the last residue at the C-terminus of the peptide. In some embodiments of the disulfide bond cyclic peptide, the first amino acid next to the N-terminus of the core motif (DXED) is cysteine, which forms a disulfide bond with the cysteine ​​located at the C-terminus of the core motif (DXED). Preferably, the two amino acids forming the ring are separated by no less than 5 amino acids, preferably, 5 or 6 amino acids. More preferably, the C-terminal cysteine ​​forming the ring is the last residue at the C-terminus of the peptide. In some embodiments, the peptide of the present invention is preferably a cyclic peptide formed by a disulfide bond and preferably consists of 8 amino acids. More preferably, the N-terminal amino group of the cyclic peptide is acetylated and / or the C-terminal carboxyl group is amidated.

[0074] The peptides of the present invention may comprise chemical modifications, for example, N-terminal acetylation, C-terminal amidation, PEG modification, lipid modification, D-amino acid substitution or non-natural amino acid substitution. In some embodiments, the peptides of the present invention are modified by covalent attachment to molecules that allow the peptide to retain its TGF-β1 activation inhibitory ability, including, for example, glycosylation, acetylation, PEGylation, phosphorylation, amidation or derivatization with known protection / blocking groups. In one embodiment, the modification is N-terminal acylation (especially acetylation). In one embodiment, the modification is C-terminal amidation. Preferably, the peptides of the present invention are linear peptides or cyclic peptides with N-terminal acylation (especially acetylation), and more preferably, the peptides also have C-terminal amidation. In some embodiments, the peptides can be attached to a biomolecule or a material for binding, labeling or identification.

[0075] The TGF-β1 activation inhibitory peptide according to the present invention is capable of inhibiting TSP-1-dependent TGF-β1 activation, and preferably has at least one of the following properties:

[0076] - Reduce the amount of active TGF-β1 in fibrotic tissue;

[0077] -Inhibits TGF-β1-mediated downstream signaling;

[0078] - Reduce collagen deposition in fibrotic tissue;

[0079] - Inhibit inflammatory and / or fibrotic lesions mediated by TGF-β1;

[0080] - Inhibits the expression of extracellular matrix-related genes stimulated by TGF-β1;

[0081] -Inhibit tumor cell migration;

[0082] -Prevention or treatment of TGF-β1 related diseases, especially fibrosis or cancer. A person skilled in the art can determine the above properties of the peptides of the present invention according to methods known in the art or described in the Examples.

[0083] In some embodiments, the peptides of the present invention comprise or consist of an amino acid sequence corresponding to one of SEQ ID NOs: 1-49. In some embodiments, the peptides of the present invention comprise or consist of one of SEQ ID NOs: 1-49. In some embodiments, the peptides of the present invention comprise or consist of an amino acid sequence corresponding to one of SEQ ID NOs: 6, 14-15, 22-26, 33, 36, 40, 43-46, and 48-49. In some embodiments, the peptides of the present invention comprise or consist of one of SEQ ID NOs: 6, 14-15, 22-26, 33, 36, 40, 43-46, and 48-49. Preferably, the peptides of the present invention block the binding of TSP-1 to its receptor in an assay as described in Example 2. In some embodiments, the peptides of the present invention exhibit a blocking rate greater than 15%, 20%, 25%, 30%, 35%, 40%, or more in an assay as described in Example 2.

[0084] In some embodiments, the peptides of the present invention comprise or consist of an amino acid sequence corresponding to one of SEQ ID NOs: 3-6, 14, 17, 22, 24-26, 31, 33, 36, 40-47. In some embodiments, the peptides of the present invention comprise or consist of an amino acid sequence corresponding to one of SEQ ID NOs: 3-6, 14, 17, 22, 24-26, 31, 33, 36, 40-47. In some embodiments, the peptides of the present invention comprise or consist of an amino acid sequence corresponding to one of SEQ ID NOs: 3, 6, 14, 22, 24-26, 33, 41-46. In some embodiments, the peptides of the present invention comprise or consist of one of SEQ ID NOs: 3, 6, 14, 22, 24-26, 33, 41-46. Preferably, the peptides of the present invention inhibit TSP-1-dependent TGF-β1 activation in an assay as described in Example 3, and preferably reduce TGF-β1 activation by 25%, 30%, 35% or more relative to a negative control without added polypeptide.

[0085] In some embodiments, the peptides of the present invention comprise or consist of an amino acid sequence corresponding to one of SEQ ID NOs: 6, 22, 25-26, 33, 40, 43, 44, 46. In some embodiments, the peptides of the present invention comprise or consist of SEQ ID NOs: 6, 22, 25-26, 33, 40, 43, 44, 46. In some embodiments, the peptides of the present invention comprise or consist of SEQ ID NO: 6. In some embodiments, the peptides of the present invention comprise or consist of SEQ ID NO: 22. In some embodiments, the peptides of the present invention comprise or consist of SEQ ID NO: 25. In some embodiments, the peptides of the present invention comprise or consist of SEQ ID NO: 26. In some embodiments, the peptides of the present invention comprise or consist of SEQ ID NO: 33. In some embodiments, the peptides of the present invention comprise or consist of SEQ ID NO: 43.

[0086] The following are some embodiments of the TGF-β1 activation inhibitory peptide of the present invention.

[0087] 1. A peptide having an amino acid sequence of the following general formula (I) or a pharmaceutically acceptable salt, solvate or prodrug thereof: 1 -X 1 -X 2 -X 3 -X 4 -X 5 -X 6 -X 7 -R 2 (I),

[0088] in,

[0089] R 1 is acetyl or does not exist;

[0090] X 1 Absent, or natural or non-natural amino acid residues;

[0091] X 2 and X 4 Each is independently a natural or non-natural amino acid residue;

[0092] X 3 It is D;

[0093] X 5 It is E;

[0094] X 6 It is D;

[0095] X 7 is a sequence of 0-7 amino acids; and

[0096] R 2 It is amino or does not exist.

[0097] 2. The peptide according to embodiment 1 or a pharmaceutically acceptable salt, solvate or prodrug thereof, wherein

[0098] X 1 absent, or selected from homoserine, allothreonine, T and S;

[0099] X 2 Selected from natural amino acids or non-natural amino acids, preferably, selected from polar side chain amino acids, such as uncharged polar side chain amino acids S, C, G, N, Q, T, Y or negatively charged polar side chain amino acids D and E, more preferably selected from amino acids N, C, Q, S, T, E and D; more preferably selected from amino acids Q, C, N, E or S;

[0100] X 3 It is D;

[0101] X 4 is selected from amino acids with small side chains, such as A, C, G, P, S, T and V, more preferably selected from amino acids A, P and S, and more preferably selected from amino acids A and P;

[0102] X 5 It is E;

[0103] X 6 It is D;

[0104] X 7 does not exist, or is selected from the amino acid sequence Z 1 Z 2 Z 3 SZ 4 LQ is a sequence of 1, 2, 3, 4, 5, 6 or 7 amino acids from the amino terminus to the carboxyl terminus.

[0105] Among them, Z 1 ,Z 2 ,Z 3 ,Z 4 independently of one another selected from natural amino acids and unnatural amino acids, preferably:

[0106] where Z 1 is the amino acid N, Q, P, A, L, V, M, or I, preferably N, P, or L, most preferably N or P;

[0107] Z 2 is an amino acid T, S, V, C, A, K, or R, preferably T, C or K, more preferably, when Z 2 When it is amino acid C or K, Z 2 With X2 or X 3 The amino acid side chains form covalent bonds,

[0108] Z 3 is amino acid A, V, L, or I, preferably A or V,

[0109] Z 4 is amino acid F, Y, H, or W, preferably F or H;

[0110] Optionally, when X 4 When A, at X 3 and X 4 0-1 small side chain amino acids, such as P or G, are inserted between, preferably, P is inserted;

[0111] Optionally, when X 7 When the peptide is composed of 1-5 amino acids, the peptide is 7 The residues may be further substituted with 0-2 additional amino acid residues, for example, 1 or 2 amino acid residues independently selected from Q, E, D and N, or 1 amino acid residue selected from C or K.

[0112] 3. The peptide according to embodiment 1-2, or a pharmaceutically acceptable salt, solvate or prodrug thereof, wherein X 1 Does not exist.

[0113] 4. The peptide according to embodiment 1-2, or a pharmaceutically acceptable salt, solvate or prodrug thereof, wherein X 1 It is T or S.

[0114] 5. The peptide according to embodiment 1-2, or a pharmaceutically acceptable salt, solvate or prodrug thereof, wherein X 1 It’s T.

[0115] 6. The peptide according to embodiments 1-5, or a pharmaceutically acceptable salt, solvate or prodrug thereof, wherein X 2 Selected from polar side chain amino acids.

[0116] 7. The peptide according to embodiments 1-5, or a pharmaceutically acceptable salt, solvate or prodrug thereof, wherein X 2 Selected from the group consisting of uncharged polar side chain amino acids S, C, G, N, Q, T and Y.

[0117] 8. The peptide according to embodiments 1-5, or a pharmaceutically acceptable salt, solvate or prodrug thereof, wherein X 2 Selected from amino acids D and E with negatively charged polar side chains.

[0118] 9. The peptide according to embodiments 1-5, or a pharmaceutically acceptable salt, solvate or prodrug thereof, wherein X 2Selected from the group consisting of amino acids N, C, homocysteine, Q, S, homoserine, T, allothreonine, E and D.

[0119] 10. The peptide according to embodiments 1-5, or a pharmaceutically acceptable salt, solvate or prodrug thereof, wherein X 2 Selected from amino acids Q, C, N, E or S.

[0120] 11. The peptide according to embodiments 1-5, or a pharmaceutically acceptable salt, solvate or prodrug thereof, wherein X 2 For Q.

[0121] 12. The peptide according to embodiments 1-5, or a pharmaceutically acceptable salt, solvate or prodrug thereof, wherein X 2 For C.

[0122] 13. The peptide according to embodiments 1-5, or a pharmaceutically acceptable salt, solvate or prodrug thereof, wherein X 2 The side chain is included in X 7 The side chains of the amino acids in the form a covalent bond, preferably, X 2 and Z 2 Both are C and their side chain sulfhydryl groups form a disulfide bond.

[0123] 14. The peptide according to embodiments 1-13, or a pharmaceutically acceptable salt, solvate or prodrug thereof, wherein X 4 Small side chain amino acids such as A, C, G, P, S, T and V.

[0124] 15. The peptide according to embodiments 1-13, or a pharmaceutically acceptable salt, solvate or prodrug thereof, wherein X 4 Selected from amino acids A, P and S.

[0125] 16. The peptide according to embodiments 1-13, or a pharmaceutically acceptable salt, solvate or prodrug thereof, wherein X 4 It is A.

[0126] 17. The peptide according to embodiments 1-13, or a pharmaceutically acceptable salt, solvate or prodrug thereof, wherein X 4 For P.

[0127] 18. The peptide according to embodiments 1-17, or a pharmaceutically acceptable salt, solvate or prodrug thereof, wherein X 7 Does not exist.

[0128] 19. The peptide according to embodiments 1-17, or a pharmaceutically acceptable salt, solvate or prodrug thereof, wherein X 7 is selected from the amino acid sequence Z 1 Z 2 Z3 SZ 4 LQ is a sequence of 1, 2, 3, 4, 5, 6 or 7 amino acids from the amino terminus to the carboxyl terminus.

[0129] 20. The peptide according to embodiments 1-17, or a pharmaceutically acceptable salt, solvate or prodrug thereof, wherein X 7 is selected from the amino acid sequence Z 1 Z 2 Z 3 SZ 4 A sequence of 1, 2, 3, 4 or 5 amino acids from the amino terminus to the carboxyl terminus.

[0130] 21. The peptide according to embodiments 1-17, or a pharmaceutically acceptable salt, solvate or prodrug thereof, wherein X 7 By Z 1 composition.

[0131] 22. The peptide according to embodiments 1-17, or a pharmaceutically acceptable salt, solvate or prodrug thereof, wherein X 7 By Z 1 Z 2 composition.

[0132] 23. The peptide according to embodiments 1-17, or a pharmaceutically acceptable salt, solvate or prodrug thereof, wherein X 7 By Z 1 Z 2 Z 3 composition.

[0133] 24. The peptide according to embodiments 1-17, or a pharmaceutically acceptable salt, solvate or prodrug thereof, wherein X 7 By Z 1 Z 2 Z 3 S composition.

[0134] 25. The peptide according to embodiments 1-17, or a pharmaceutically acceptable salt, solvate or prodrug thereof, wherein X 7 By Z 1 Z 2 Z 3 SZ 4 composition.

[0135] 26. The peptide according to embodiments 1-17, or a pharmaceutically acceptable salt, solvate or prodrug thereof, wherein X 7 By Z 1 Z 2 Z 3 SZ 4 L composition.

[0136] 27. The peptide according to embodiments 1-17, or a pharmaceutically acceptable salt, solvate or prodrug thereof, wherein X 7 By Z 1 Z 2 Z 3 SZ 4 LQ composition.

[0137] 28. The peptide according to embodiments 1-27, or a pharmaceutically acceptable salt, solvate or prodrug thereof, wherein Z 1 is N, Q, P, A, N-methylalanine, L, norleucine, V, M, N-methylmethionine, or I.

[0138] 29. The peptide according to embodiments 1-27, or a pharmaceutically acceptable salt, solvate or prodrug thereof, wherein Z 1 It's N.

[0139] 30. The peptide according to embodiments 1-27, or a pharmaceutically acceptable salt, solvate or prodrug thereof, wherein Z 1 It’s P.

[0140] 31. The peptide according to embodiments 1-27, or a pharmaceutically acceptable salt, solvate or prodrug thereof, wherein Z 1 It’s L.

[0141] 32. The peptide according to embodiments 1-31, or a pharmaceutically acceptable salt, solvate or prodrug thereof, wherein Z 2 is T, allothreonine, S, homoserine, V, C, homocysteine, A, N-methylalanine, K, or R.

[0142] 33. The peptide according to embodiments 1-31 or a pharmaceutically acceptable salt, solvate or prodrug thereof, wherein Z 2 It’s T.

[0143] 34. The peptide according to embodiments 1-31 or a pharmaceutically acceptable salt, solvate or prodrug thereof, wherein Z 2 is C, and with X 2 Covalently linked via side chains.

[0144] 35. The peptide according to embodiments 1-31, or a pharmaceutically acceptable salt, solvate or prodrug thereof, wherein Z 2 is K, and with X 3 Covalently linked via side chains.

[0145] 36. The peptide according to embodiments 1-35, or a pharmaceutically acceptable salt, solvate or prodrug thereof, wherein Z 3 is A, N-methylalanine, V, L, norleucine, or I.

[0146] 37. The peptide according to embodiments 1-35, or a pharmaceutically acceptable salt, solvate or prodrug thereof, wherein Z 3 It's A, especially Z 1 When it is N or P.

[0147] 38. The peptide according to embodiments 1-35, or a pharmaceutically acceptable salt, solvate or prodrug thereof, wherein Z 3 It’s V.

[0148] 39. The peptide according to embodiments 1-38, or a pharmaceutically acceptable salt, solvate or prodrug thereof, wherein Z 4 It is an aromatic amino acid.

[0149] 40. The peptide according to embodiments 1-38, or a pharmaceutically acceptable salt, solvate or prodrug thereof, wherein Z 4 is F, N-methylphenylalanine, homophenylalanine, Y, H, or W.

[0150] 41. The peptide according to embodiments 1-38, or a pharmaceutically acceptable salt, solvate or prodrug thereof, wherein Z 4 It is F or H.

[0151] 42. The peptide according to embodiments 1-38, or a pharmaceutically acceptable salt, solvate or prodrug thereof, wherein Z 4 It’s H.

[0152] 43. The peptide according to embodiments 1-42, or a pharmaceutically acceptable salt, solvate or prodrug thereof, wherein X 7 By Z 1 Composed of; or consisting of Z 1 Z 2 Composed of; or consisting of Z 1 Z 2 Z 3 Composed of; or consisting of Z 1 Z 2 Z 3 S; or Z 1 Z 2 Z 3 SZ 4 composition.

[0153] 44. The peptide according to embodiments 1-43, or a pharmaceutically acceptable salt, solvate or prodrug thereof, wherein X 7 It is an amino acid sequence selected from the group consisting of: N; P; L; NTA; NTV; PTA; NTAS; PTAS; NTVSF; NTASF; NTASH; NTVSFQ.

[0154] 45. The peptide according to embodiments 1-44, or a pharmaceutically acceptable salt, solvate or prodrug thereof, wherein the peptide is 7 Afterwards, 0-2 additional amino acid residues are added.

[0155] 46. ​​The peptide according to embodiments 1-45, or a pharmaceutically acceptable salt, solvate or prodrug thereof, wherein the peptide comprises an amino acid sequence selected from the group consisting of: DAED; DPED; DAEDN; DPEDN; or DAEDP.

[0156] 47. The peptide according to embodiments 1-46, or a pharmaceutically acceptable salt, solvate or prodrug thereof, wherein X 1 is T and X 2 For Q.

[0157] 48. The peptide according to embodiments 1-47, or a pharmaceutically acceptable salt, solvate or prodrug thereof, wherein R 1 It is acetyl.

[0158] 49. The peptide according to embodiments 1-48, or a pharmaceutically acceptable salt, solvate or prodrug thereof, wherein R 2 It is an amino group.

[0159] 50. An amino acid sequence peptide having the following general formula (II) or a pharmaceutically acceptable salt, solvate or prodrug thereof: R 1 -X 1 -X 2 -DX 4 -EDX 7 -R 2 (II),

[0160] in,

[0161] R 1 is acetyl or does not exist;

[0162] R 2 It is amino or does not exist;

[0163] X 1 is T or S;

[0164] X 2 selected from polar side chain amino acids;

[0165] X 4 Selected from amino acids A or P;

[0166] X 7 is selected from the amino acid sequence Z 1 TZ 3 SZ 4A sequence of 1, 2, 3, 4 or 5 amino acids from the amino terminus to the carboxyl terminus, wherein

[0167] where Z 1 is N, P or L;

[0168] Z 3 is A, V, L, or I,

[0169] Z 4 It is F, Y, H, or W, preferably F or H.

[0170] 51. The peptide according to embodiment 50, or a pharmaceutically acceptable salt, solvate or prodrug thereof, wherein X 2 is selected from the group consisting of amino acids Q, N, C, S, T, E and D; preferably selected from the group consisting of Q, N, E or S; more preferably X 2 It is Q or N.

[0171] 52. The peptide according to embodiment 50, or a pharmaceutically acceptable salt, solvate or prodrug thereof, wherein X 2 It's Q.

[0172] 53. The peptide according to embodiments 50-52, or a pharmaceutically acceptable salt, solvate or prodrug thereof, wherein Z 1 is N and Z 3 A or V; or Z 1 Is P and Z 3 It’s A.

[0173] 54. The peptide according to embodiments 50-53, or a pharmaceutically acceptable salt, solvate or prodrug thereof, wherein Z 4 It is F or H.

[0174] 55. An amino acid sequence peptide having the following general formula (III) or a pharmaceutically acceptable salt, solvate or prodrug thereof: R 1 -X 1 -Ring(-CDX 4 -EDZ 1 -C)-X 8 -R 2 (III),

[0175] in,

[0176] R 1 is acetyl or does not exist;

[0177] R 2 It is amino or does not exist;

[0178] X 1 is T or S;

[0179] X4 is selected from amino acids with small side chains, such as A, C, G, P, S, T and V, more preferably selected from amino acids A, P and S, and more preferably selected from amino acids A and P;

[0180] Z 1 is a natural amino acid or an unnatural amino acid;

[0181] X 8 Absent, or a sequence of 1-5 amino acids.

[0182] 56. The peptide according to embodiment 55, or a pharmaceutically acceptable salt, solvate or prodrug thereof, wherein X 4 It is A, C, G, P, S, T or V.

[0183] 57. The peptide according to embodiment 55, or a pharmaceutically acceptable salt, solvate or prodrug thereof, wherein X 4 It’s A.

[0184] 58. The peptide according to embodiment 55, or a pharmaceutically acceptable salt, solvate or prodrug thereof, wherein X 4 It’s P.

[0185] 59. The peptide according to embodiments 55-58, or a pharmaceutically acceptable salt, solvate or prodrug thereof, wherein Z 1 It is a polar uncharged amino acid or a nonpolar amino acid.

[0186] 60. The peptide according to embodiments 55-58, or a pharmaceutically acceptable salt, solvate or prodrug thereof, wherein Z 1 It is N, Q, P, A, L, V, M, or I.

[0187] 61. The peptide according to embodiments 55-58, or a pharmaceutically acceptable salt, solvate or prodrug thereof, wherein Z 1 It's N.

[0188] 62. The peptide according to embodiments 55-61, or a pharmaceutically acceptable salt, solvate or prodrug thereof, wherein X 8 Does not exist.

[0189] 63. The peptide according to embodiments 55-61, or a pharmaceutically acceptable salt, solvate or prodrug thereof, wherein X 8 Selected from the amino acid sequence Z 3 SZ 4 LQ is a sequence of 1, 2, 3, 4 or 5 amino acids from the amino terminus to the carboxyl terminus, wherein Z 3 is A, V, L, or I, and Z 4 It is F, Y, H, or W, preferably F or H.

[0190] 64. The peptide according to embodiment 63, or a pharmaceutically acceptable salt, solvate or prodrug thereof, wherein Z 3 is A or V, and Z 4 It is F or H.

[0191] 65. The peptide according to embodiments 55-64, or a pharmaceutically acceptable salt, solvate or prodrug thereof, wherein the peptide consists of 5 to 13 amino acids, preferably 7, 8, 9, 10 or 11 amino acids.

[0192] 66. The peptide according to embodiments 1-54, or a pharmaceutically acceptable salt, solvate or prodrug thereof, wherein the peptide consists of 5 to 13 amino acids, preferably 7, 8, 9, 10 or 11 amino acids.

[0193] 67. The peptide according to embodiment 66, or a pharmaceutically acceptable salt, solvate or prodrug thereof, wherein the peptide is a linear peptide.

[0194] 68. The peptide according to embodiment 66, or a pharmaceutically acceptable salt, solvate or prodrug thereof, wherein the peptide is a cyclic peptide, and preferably, the cyclization region of the peptide comprises a core motif (DXED).

[0195] 69. The peptide according to embodiment 68, or a pharmaceutically acceptable salt, solvate or prodrug thereof, wherein the peptide is head-to-tail cyclic or side-chain cyclic.

[0196] 70. The peptide according to embodiment 68, or a pharmaceutically acceptable salt, solvate or prodrug thereof, wherein the peptide is head-to-tail cyclic.

[0197] 71. The peptide according to embodiment 68, or a pharmaceutically acceptable salt, solvate or prodrug thereof, wherein the peptide has a side chain that forms a ring.

[0198] 72. The peptide according to embodiment 68, or a pharmaceutically acceptable salt, solvate or prodrug thereof, wherein the peptide is coupled to form a ring via the side chains of amino acids at positions 3 and 8, or via the side chains of amino acids at positions 2 and 8.

[0199] 73. The peptide according to embodiments 71-72, or a pharmaceutically acceptable salt, solvate or prodrug thereof, wherein the peptide is cyclized via a lactam bond formed between amino acid side chains.

[0200] 74. The peptide according to embodiments 71-72, or a pharmaceutically acceptable salt, solvate or prodrug thereof, wherein the peptide is cyclized via disulfide bonds formed between amino acid side chains.

[0201] 75. The peptide according to embodiments 71-74, or a pharmaceutically acceptable salt, solvate or prodrug thereof, wherein the peptide is cyclized through the side chain,

[0202] Among them, X 3 It's D, Z 2 is K, and the peptide is 3 With Z 2 cyclization of the lactam bond formed between the side chains of

[0203] Among them, X 2 It's C, Z 2 is C, and the peptide is connected by X 2 With Z 2 The disulfide bonds formed between the side chains of

[0204] Preferably, when Z 2 When it is amino acid C or K, X 7 By Z 1 Z 2 Composition, and X 1 T or S.

[0205] 76. A peptide or a pharmaceutically acceptable salt, solvate or prodrug thereof, wherein the peptide has an amino acid sequence corresponding to any one selected from SEQ ID NOs: 1 to 49, or comprises or consists of any one selected from SEQ ID NOs: 1 to 49.

[0206] 77. A peptide or a pharmaceutically acceptable salt, solvate or prodrug thereof, wherein the peptide has an amino acid sequence corresponding to any one selected from SEQ ID NO: 6, 22, 25, 26, 33, 40, 43, 44 and 46, or comprises any one selected from SEQ ID NO: 6, 22, 25, 26, 33, 40, 43, 44 and 46.

[0207] 78. A peptide or a pharmaceutically acceptable salt, solvate or prodrug thereof, wherein the peptide comprises or consists of SEQ ID NO: 6.

[0208] 79. A peptide or a pharmaceutically acceptable salt, solvate or prodrug thereof, wherein the peptide comprises or consists of SEQ ID NO: 22.

[0209] 80. A peptide or a pharmaceutically acceptable salt, solvate or prodrug thereof, wherein the peptide comprises or consists of SEQ ID NO: 25.

[0210] 81. A peptide or a pharmaceutically acceptable salt, solvate or prodrug thereof, wherein the peptide comprises or consists of SEQ ID NO: 26.

[0211] 82. A peptide or a pharmaceutically acceptable salt, solvate or prodrug thereof, wherein the peptide comprises or consists of SEQ ID NO: 33.

[0212] 83. A peptide or a pharmaceutically acceptable salt, solvate or prodrug thereof, wherein the peptide comprises or consists of SEQ ID NO: 40.

[0213] 84. A peptide or a pharmaceutically acceptable salt, solvate or prodrug thereof, wherein the peptide comprises or consists of SEQ ID NO: 43.

[0214] 85. A peptide or a pharmaceutically acceptable salt, solvate or prodrug thereof, wherein the peptide comprises or consists of SEQ ID NO: 44.

[0215] 86. A peptide or a pharmaceutically acceptable salt, solvate or prodrug thereof, wherein the peptide comprises or consists of SEQ ID NO: 46.

[0216] 87. A peptide or a pharmaceutically acceptable salt, solvate or prodrug thereof, wherein the peptide has 1, 2, 3, 4 or 5 amino acid differences compared to the peptide according to one of embodiments 76-86, preferably 1 or 2 amino acid substitutions, deletions or additions, preferably, the substitutions occur in amino acid residues outside the core motif (DXED), such as conservative amino acid substitutions.

[0217] 88. The peptide according to embodiments 1-87, or a pharmaceutically acceptable salt, solvate or prodrug thereof, wherein the peptide is chemically modified.

[0218] 89. The peptide according to embodiments 1-88, or a pharmaceutically acceptable salt, solvate or prodrug thereof, wherein the peptide comprises a PEG modification.

[0219] 90. The peptide according to embodiments 1-89, or a pharmaceutically acceptable salt, solvate or prodrug thereof, wherein the peptide comprises a lipid modification.

[0220] 91. The peptide according to embodiments 1-90, or a pharmaceutically acceptable salt, solvate or prodrug thereof, wherein the peptide comprises D-amino acid substitutions and / or non-natural amino acid substitutions.

[0221] Preparation method of the inhibitory peptide of the present invention

[0222] In one aspect, the present invention provides a method for producing the inhibitory peptide of the present invention. The peptide of the present invention can be produced using chemical synthesis, fermentation, or genetic recombination technology.

[0223] Methods for chemically synthesizing peptides are well known in the art and include solid-phase synthesis and liquid-phase synthesis techniques. The desired peptide chain can be synthesized using the solid-phase peptide synthesis (SPPS) technique developed by Merrifield. An automated peptide synthesizer can also be used for synthesis. Subsequent purification of SPPS peptide products is simpler and easier than recombinantly synthesized peptides. Therefore, it is preferred to use SPPS technology to synthesize the short peptides of the present invention.

[0224] SPPS technology generally consists of coupling and deprotection cycles. In the Fmoc synthesis strategy, in the coupling step, a condensing agent such as HBTU / HATU / DIC is used to couple the Fmoc-protected amino acid (Fmoc-AA-OH) to a solid polymer resin; in the deprotection step, the protecting group Fmoc is removed from the amino acid by a deprotecting agent such as piperidine to release the amino group, thereby entering the next coupling and deprotection cycle. The entire process can be carried out in a sieve reactor until the final peptide is synthesized and cut from the resin. Afterwards, the synthesized peptide product can be purified and detected by high-performance liquid chromatography.

[0225] There are various resins, such as 4-toluenehydrogenamine (HMBA) resin, Wang resin, 2-chlorotrityl chloride (CTC) resin and Merrifield resin, which can be used for solid phase synthesis of peptides. In addition, a variety of functional resins have been developed by coupling the resin with different linkers to allow peptide cyclization in the solid phase.

[0226] In one embodiment, the present invention provides a method for preparing the peptide of the present invention, comprising: (1) synthesizing a linear peptide having a specified sequence, for example, using 4-toluenehydrogenamine (HMBA) resin as a starting material, Fmoc-protected amino acids as monomers, hexahydropyridine / DMF solution as a deprotection reagent, and sequentially connecting amino acids one by one under the action of a condensing agent and alkaline conditions to produce the linear peptide; (2) optionally, cyclizing the linear peptide obtained in step (1); and (3) purifying the peptide product obtained in step (1) or (2).

[0227] In another embodiment, the present invention provides a method for preparing a peptide of the present invention, comprising: (1) recombinantly expressing a linear peptide having a specified sequence; (2) optionally, cyclizing the linear peptide obtained in step (1); and (3) purifying the peptide product obtained in step (1) or (2). In some aspects, the present invention also provides a nucleic acid comprising an amino acid sequence encoding a peptide of the present invention, a vector (e.g., an expression vector), and a host cell comprising the nucleic acid.

[0228] Peptide cyclization is a common peptide modification technique that can include various strategies, such as head-to-tail cyclization, side-chain-to-side-chain cyclization, and main-chain-to-side-chain cyclization. Typically, a single linear peptide is too flexible without being linked to other peptides. Cyclization, however, can promote the formation of secondary peptide structure by organizing intramolecular interactions, thereby improving peptide stability.

[0229] In one embodiment according to the present invention, the peptide of the present invention forms a single cyclic peptide by head-to-tail cyclization. In another embodiment according to the present invention, the peptide of the present invention forms a lactam bridge between the side chains of glutamic acid (E) or aspartic acid (D) and lysine (K) through side chain cyclization. In another embodiment according to the present invention, the peptide of the present invention forms a disulfide bond between the side chains of two cysteine ​​(C) residues through side chain cyclization. In the embodiment of the cyclic peptide of the present invention comprising a lactam bridge, preferably, the lysine amino acid residue is located at the C-terminus of the core sequence (DXED) of the present invention and forms the lactam bridge with the first aspartic acid D residue of the core sequence (DXED). More preferably, the K residue and the D residue are separated by at least 4 residues, for example, 4, 5, or 6 residues, and preferably the lysine residue is the last residue at the C-terminus of the peptide of the present invention. In the embodiment of the cyclic peptide of the present invention comprising a disulfide bond, preferably, the two cysteine ​​residues used for cyclization are located at the N-terminus and C-terminus of the core sequence (DXED), respectively. More preferably, the two cysteine ​​residues are separated by at least 5 residues, for example, 5, 6, 7, or 8 residues. In a preferred embodiment, the third amino acid of the peptide of the present invention is D and the eighth amino acid is K, and the side chains of the peptide of the present invention form a lactam bond to form a ring. In another preferred embodiment, the second amino acid of the peptide of the present invention is C and the eighth amino acid is C, and the side chains of the peptide of the present invention form a disulfide bond to form a ring.

[0230] After peptide synthesis, it can be modified with or without the use of medical chemistry techniques. In some cases, it may be advantageous to modify the peptide, for example, by modifying it to simulate, stabilize or construct a more suitable secondary structure to improve the biological activity of the peptide drug, and / or improve the selectivity, stability and solubility of the peptide drug. Peptide modifications can be amino acid substitutions or residue modifications of non-essential amino acids, and / or modifications of the N-terminus and / or C-terminus of the peptide, such as N-terminal acetylation modification and C-terminal amidation modification. Modifications can also be substitutions of key amino acid residues that affect the biological activity of the peptide to seek changes in activity.

[0231] In some cases, the peptide may comprise non-natural amino acids. Depending on the needs, the peptide may comprise 1, 2, 3, 4, 5, or 6 or more non-natural amino acids. Alternatively, the amino acids comprising the peptide may each be independently selected from natural amino acids.

[0232] Chemically synthesized peptides often carry free amino and free carboxyl groups. In some cases, the termini of the peptides can be blocked, i.e., N-terminal acetylation and C-terminal amidation, to make the synthetic peptides closer to natural protein mimics and improve the stability of the peptides.

[0233] Thus, in some aspects, the peptides of the invention can comprise D-amino acids, unnatural amino acids, amino acid analogs, and / or group substitutions and modifications; or can be linked to a polymer or to a drug carrier.

[0234] Pharmaceutical composition

[0235] In one aspect, the present invention also provides a pharmaceutical composition comprising a peptide of the present invention or a pharmaceutically acceptable salt, solvate or prodrug thereof. As is known in the art, the pharmaceutical composition may also optionally comprise suitable pharmaceutical excipients, pharmaceutical carriers, pharmaceutical excipients, including buffers.

[0236] The administration routes for peptide administration include, but are not limited to, subcutaneous, intramuscular and intravenous administration, mucosal administration (e.g., nasal administration, pulmonary mucosal administration, sublingual administration), oral administration (e.g., with the addition of a gastrointestinal permeation enhancer or carrier), and transdermal administration. The peptide can be formulated into any form suitable for administration, such as an injection, e.g., an intravenous injection or intravenous drip, a lyophilized powder, etc. Excipients for different formulations are known in the art.

[0237] As is known in the art, pharmaceutical compositions may also contain other therapeutic agents that are beneficial for the specific disease to be treated. In some embodiments, the present invention also provides pharmaceutical compositions comprising a peptide of the present invention or a pharmaceutically acceptable salt, solvate, or prodrug thereof, and further comprising other therapeutic agents that are beneficial for the specific disease to be treated. In such embodiments, the peptide of the present invention may be contained in the same or different compositions as the other therapeutic agent; and may be administered simultaneously, sequentially, or in any order and in any dosing regimen with the other therapeutic agent.

[0238] In some embodiments, the pharmaceutical composition of the present invention may contain, in particular, the peptide of the present invention according to the aforementioned embodiments 1-91 or a pharmaceutically acceptable salt thereof, in particular, the peptide of the present invention according to one of the aforementioned embodiments 76-86 or a pharmaceutically acceptable salt thereof, in particular, the peptide of the present invention according to one of the aforementioned embodiments 78-86 or a pharmaceutically acceptable salt thereof.

[0239] Treatment methods and uses

[0240] In one aspect, the present invention provides a peptide of the present invention or a pharmaceutically acceptable salt, solvate or prodrug thereof, or a pharmaceutical composition of the present invention for use as a medicament for preventing or treating a TGF-β-related disease in a subject, and corresponding disease treatment and prevention methods.

[0241] The term "TGF-β-related diseases" herein refers to diseases and conditions associated with and / or caused by TSP-1-involved TGF-β1 activation or pathologically increased TGF-β1 activity, including but not limited to fibrosis, chronic inflammation, and cancer. Such TGF-β-related diseases would benefit from inhibition of TSP-1-involved TGF-β1 activation.

[0242] As used herein, the term "subject" or "patient" or "individual" includes any human or non-human animal. The term "non-human animal" includes all vertebrates, e.g., mammals and non-mammals, such as non-human primates, sheep, dogs, cats, horses, cows, chickens, amphibians, reptiles, etc. Preferably, the subject of the present invention is a human.

[0243] As used herein, the term "effective amount" or "effective dose" refers to an amount of a peptide of the present invention or a pharmaceutically acceptable salt, solvate or prodrug thereof, which, when administered alone or in combination with other therapeutic agents to a cell, tissue or subject, effectively prevents or improves the symptoms of one or more diseases or conditions or the development of the disease or condition. An effective dose also refers to an amount sufficient to cause symptomatic improvement, such as an amount that treats, cures, prevents or improves a related medical condition or increases the rate of treatment, cure, prevention or improvement of such a condition. When a single active ingredient is administered to an individual, the effective dose refers only to that ingredient. When administered in combination, the effective dose refers to the combined amount of active ingredients that cause a therapeutic or preventive effect, whether in combination, administered sequentially or administered simultaneously. In some embodiments, the effective amount will result in at least 10%, typically at least 20%, preferably at least about 30%, more preferably at least 40%, and most preferably at least 50% improvement in a diagnostic criterion or parameter.

[0244] In some embodiments according to the present invention, " treatment " of disease or condition refers to improving disease or condition (that is, slowing down or preventing or reducing at least one of the progress of disease or its clinical symptoms). In other embodiments, " treatment " refers to alleviating or improving at least one physical parameter, including those physiological parameters that may not be discerned by the patient. In other embodiments, " treatment " refers to regulating disease or condition in both aspects, for example, physically (for example, the stabilization of discernible symptoms), physiologically (for example, the stabilization of physical parameters). Unless clearly described in this article, the method for the treatment and / or prevention of the disease for assessment is generally known in the art.

[0245] In further embodiments according to the present invention, "prevention" of a disease or condition includes inhibition of the onset or development of the disease or condition or symptoms of a particular disease or condition. Generally, in the context of fibrosis or cancer, the term "prevention" refers to the administration of a drug before the onset of signs or symptoms of fibrosis or cancer, particularly in a subject at risk of developing the disease or condition.

[0246] Therefore, in some embodiments, the present invention provides a method for preventing or treating a TGF-β-related disease, comprising administering to a subject in need thereof an effective amount of a peptide according to the present invention or a pharmaceutically acceptable salt, solvate or prodrug thereof, or a pharmaceutical composition according to the present invention. Optionally, the method further comprises administering to the subject an effective amount of a second therapeutic agent, such as a chemotherapeutic agent or an immunotherapeutic agent. Preferably, the TGF-β-related disease treated according to the method of the present invention has macrophage infiltration of the lesion site. In some embodiments, the TGF-β-related disease is associated with tissue damage, inflammation or fibrosis. In some embodiments, the TGF-β-related disease involves metastasis of tumor cells.

[0247] In some embodiments, in the prevention or treatment method of the present invention, the peptide of the present invention or its pharmaceutically acceptable salt according to the aforementioned embodiments 1-91, especially the peptide of the present invention or its pharmaceutically acceptable salt according to one of the aforementioned embodiments 76-86, and especially the peptide of the present invention or its pharmaceutically acceptable salt according to one of the aforementioned embodiments 78-86 can be particularly used.

[0248] Treatment of fibrotic disorders

[0249] In some embodiments, the condition associated with pathological increase in TGF-β activity is fibrosis. In one embodiment, the present invention provides a peptide of the present invention or a pharmaceutically acceptable salt, solvate or prodrug thereof, or a pharmaceutical composition of the present invention for use as a medicament for preventing or treating a fibrotic condition in a subject, and corresponding treatment and prevention methods.

[0250] Currently, the known diseases characterized by fibrosis encompass a vast spectrum of diseases. These can be categorized based on the tissues and organs in which they occur, such as fibrosis of the lungs, liver, kidneys, and bone marrow; they can also be grouped within common disease categories, such as chronic inflammatory diseases, tumors, and immune disorders. The presence of fibrosis as a pathological characteristic is becoming an important basis for reclassifying, naming, diagnosing, and treating fibrotic conditions, moving beyond the limitations of the known site of disease onset or traditional disease classifications.

[0251] As used herein, "fibrotic disorder" refers to a disease characterized by a fibrotic pathological phenotype, and is not limited to a specific site of disease onset or traditional classification of the disease. Examples of fibrotic diseases treatable by the methods of the present invention include, but are not limited to, fibrosis of various tissues and organs, such as pulmonary fibrosis, liver fibrosis, and renal fibrosis; as well as chronic inflammatory, tumor, and immune diseases.

[0252] In some embodiments, therefore, the present invention provides a method for preventing or treating a fibrotic disorder in a subject, comprising administering to a subject in need thereof a prophylactic or therapeutically effective amount of a peptide according to the present invention or a pharmaceutically acceptable salt, solvate or prodrug thereof, or a pharmaceutical composition according to the present invention. In some embodiments, the fibrotic disorder is liver fibrosis, pulmonary fibrosis, kidney fibrosis, bone marrow fibrosis, skin fibrosis, or cardiac fibrosis.

[0253] In some embodiments, the fibrotic disorder is pulmonary fibrosis, eg, idiopathic pulmonary fibrosis (IPF).In some embodiments, the method comprises administering a peptide of the invention to the subject by inhalation, eg, as an aerosolized formulation.

[0254] The therapeutic uses and methods described herein may also include co-formulating and / or co-administering the peptides of the present invention or pharmaceutically acceptable salts, solvates or prodrugs thereof or pharmaceutical compositions thereof with another therapeutically effective agent to prevent and / or treat pathological fibrotic diseases.

[0255] Cancer treatment

[0256] In some embodiments, the condition associated with the pathological increase in TGF-β activity is cancer. In one embodiment, the present invention provides a peptide of the present invention or a pharmaceutically acceptable salt, solvate or prodrug or pharmaceutical composition thereof for use as a drug for preventing or treating cancer in a subject, and corresponding treatment and prevention methods. In some embodiments, the cancer is a solid tumor, preferably selected from: lung cancer, liver cancer, breast cancer, uterine cancer, prostate cancer, pancreatic cancer, colon cancer, skin cancer, central nervous system cancer, fibromyoma, fibroma, fibroadenoma and fibrosarcoma. In some embodiments, cancer is sarcoma, pancreatic cancer, glioblastoma, head and neck cancer, melanoma, breast cancer or colorectal cancer. In some embodiments, cancer is selected from squamous cell carcinoma, epidermoid carcinoma, urothelial carcinoma, adenocarcinoma, adrenocortical carcinoma, basal cell carcinoma, ductal carcinoma in situ (DCIS), invasive ductal carcinoma, thymic carcinoma and renal cell carcinoma.

[0257] In some embodiments, the method comprises administering to the subject a peptide of the present invention or a pharmaceutically acceptable salt, solvate or prodrug thereof or a pharmaceutical composition thereof by oral, intravenous, intratumoral, transdermal, subcutaneous or topical administration. In some embodiments, the method comprises contacting the subject's cancerous tissue (e.g., cancerous skin tissue) with a formulation containing a therapeutically effective amount of a peptide of the present invention or a pharmaceutically acceptable salt, solvate or prodrug thereof or a pharmaceutical composition thereof for a time sufficient to treat the cancer.

[0258] The therapeutic uses and methods described herein may also include co-formulating and / or co-administering the peptides of the present invention, or pharmaceutically acceptable salts, solvates, or prodrugs thereof, or pharmaceutical compositions thereof, with another therapeutically effective agent to prevent and / or treat cancer.

[0259] Other applications

[0260] In some aspects, the present invention also provides the following uses of the peptide of the present invention or a pharmaceutically acceptable salt, solvate or prodrug thereof:

[0261] - Blocking the binding of CD36 to TSP-1;

[0262] - Inhibits TSP-1-dependent TGF-β1 activation;

[0263] - Reduce the amount of active TGF-β1 in fibrotic tissue;

[0264] -Inhibits TGF-β1-mediated downstream signaling;

[0265] - Reduce collagen deposition in fibrotic tissue;

[0266] - Inhibit inflammatory and / or fibrotic lesions mediated by TGF-β1;

[0267] - Inhibits the expression of extracellular matrix-related genes stimulated by TGF-β1;

[0268] -Inhibit tumor cell migration;

[0269] -Prevention or treatment of TGF-β1-related diseases, especially fibrosis or cancer;

[0270] Or used for preparing medicines used in the above purposes.

[0271] The above-mentioned application can be in vitro or in vivo. In a preferred embodiment, the application is an in vivo disease therapeutic or preventive application. In the in vivo application method, the peptide of the present invention or a pharmaceutically acceptable salt, solvate or prodrug thereof is administered in a therapeutically effective amount, optionally in combination with other therapeutic agents.

[0272] For the prevention or treatment of disease, the appropriate dosage of the peptide of the present invention or its pharmaceutically acceptable salt, solvate or prodrug (when used alone or in combination with one or more other therapeutic agents) will depend on the type of disease to be treated, the type of specific drug, the severity and course of the disease, whether it is administered for preventive or therapeutic purposes, previous treatment, the patient's clinical history and response to the drug, and the judgment of the attending physician. The drug can be appropriately administered to the patient as a single treatment or over a series of treatments.

[0273] In another aspect, the present invention also provides use of the peptide of the present invention or a pharmaceutically acceptable salt, solvate or prodrug thereof in the preparation of a medicament for use in the aforementioned method (eg, for treatment).

[0274] In order to assist in understanding the present invention, the following examples are further given. However, it should be understood that these examples are not intended to and should not in any way limit the scope of protection of the present invention.

[0275] For the purposes of the present invention, when referring to amino acid residues herein, the following amino acid codes apply, including both three-letter and one-letter codes: Example

[0276] Example 1 Polypeptide Preparation

[0277] The polypeptides used in the embodiments of the present invention were prepared using the Fmoc (9-fluorenylmethyloxycarbonyl) solid-phase synthesis method. Using the carboxyl-terminal amino acid of the linear peptide as the starting point for synthesis, Rink Amide MBHA Resin (Gill Biochemical, 49101) was first soaked in a DCM solution for 30 minutes in a peptide solid-phase synthesis reaction column for resin activation. The resin was then drained and a 20% hexahydropyridine + 80% DMF solution was added. The mixture was stirred and agitated with nitrogen for 30 minutes to perform a deprotection reaction. The Fmoc-protected amino acid, condensing agent, and organic base were then added for reaction. After the reaction, the reaction was completed using the ninhydrin colorimetry method. The above steps of adding, reacting, and detecting were repeated continuously according to the amino acid sequence of each peptide (from the carboxyl terminus to the amino terminus) until the last amino acid was reached. The target peptide was then cleaved from the resin to obtain a crude peptide.

[0278] The steps for cyclizing peptides are as follows:

[0279] (1) Disulfide bond cyclization: The linear peptide synthesized and cleaved on the resin as described above is stirred in an aqueous solution at pH = 7.5-8 (peptide concentration 1 g / L) for more than 12 hours to form a ring.

[0280] (2) Side chain cyclization: During the aforementioned linear peptide synthesis process, special side chain protecting groups are selected, such as D (oall) and K (alloc), to protect the side chain carboxyl group of D and the side chain amino group of K. After the linear peptide sequence is synthesized, -oall / -alloc is removed on the resin, and then cyclization is carried out using the corresponding condensing agent.

[0281] (3) Head-to-tail cyclization: The linear peptide synthesized and cleaved on the resin as described above is prepared into fully protected peptide fragments. The cyclization is carried out by adding a corresponding condensing agent in DMF or other solvents. After the reaction is completed, the product is precipitated from water, dried, and the protecting groups on the cyclic peptide are cleaved to obtain the crude deprotected head-to-tail cyclized peptide.

[0282] The crude linear and cyclic peptides synthesized were separated and purified by HPLC using a C18 column, using eluent A of 0.1% TFA / acetonitrile and eluent B of 0.1% TFA / water. The main peak was collected, and the molecular weight of the product was consistent with the theoretical value by mass spectrometry. The refined peptides were then lyophilized. The sequences of the synthesized peptides are shown in Table 1, where each sequence number represents a peptide composed of the "Sequence" and "Modification" sections of the sequence description.

[0283] Table 1 Peptide sequence list

[0284] Example 2 In vitro blocking activity test of polypeptide targets

[0285] The ELISA method was used to detect the ability of the peptide to block the binding of TSP-1 protein to TSP-1 receptor protein CD36. TSP-1 protein (novoprotein, product number CU45) was diluted to 2 μg / mL using 100 mM carbonate buffer and added to the sample wells (100 μL / well) of a 96-well ELISA plate (Thermo, product number 437111) and incubated overnight at 4°C; after washing the plate twice with 1×PBST solution, 200 μL of 1.5% BSA (Sigma, product number B2064) was added to each well and incubated at 37°C for 2 hours; after washing the plate three times with 1×PBST solution, the analyte was added (100 μL / well, 3 parallel wells, and 0.25 μg / mL of Fc-tagged CD36 protein (R&D Systems, Catalog No. 1955-CD-050); a peptide-treated group (a mixture of Fc-tagged CD36 protein and 100 μM peptide at a final concentration of 0.25 μg / mL) was added and incubated at 37°C for 3 h. After washing the plates five times with 1× PBST, a 1:500 dilution of FITC-labeled anti-Fc tag antibody (Biyuntian, Catalog No. A0556) was added and incubated at 37°C for 1 h. After washing the plates five times with 1× PBST, the FITC fluorescence signal intensity in each well was measured using a multi-function microplate reader (TECAN, Model Infinite 200Pro). The blocking rate of the peptide's ability to block CD36 binding to TSP-1 was calculated by calculating the ratio of the fluorescence signal in the peptide-treated group to the control group (i.e., (fluorescence signal of the control group - fluorescence signal of the peptide-treated group) / fluorescence signal of the control group × 100%) to assess the peptide's ability to block CD36 binding to TSP-1 (Table 2-1). Based on the single concentration blocking rate results and the characteristics of the peptide molecules, the peptide molecules were selected for concentration (0 μM, 0.5 μM, 1 μM, 5 μM, 10 μM, 100 μM) dependent experiments to determine the IC of the peptides blocking the binding of CD36 to TSP-1 protein. 50The values ​​were used to further evaluate the ability of the preferred polypeptide molecules to block the binding of CD36 to TSP-1 (Table 2-2). The test results showed that the polypeptides of the present invention were able to inhibit the binding of CD36 to TSP-1.

[0286] Table 2-1 In vitro peptide target binding test data

[0287] Table 2-2 IC of peptides blocking the binding of CD36 to TSP-1 protein 50 value

[0288] Example 3 Inhibitory test of polypeptide on TSP-1-dependent TGF-β activation in cell model

[0289] This example uses a cell model with high TSP-1 expression to detect the inhibitory effect of the polypeptide on TSP-1-dependent TGF-β activation. The experiment uses phorbol 12-myristate 13-acetate (PMA) (Sigma, Catalog No. P1585) to induce THP-1 cells (American Type Culture Collection, ATCC, TIB-202) to differentiate into macrophage-like cells with obvious macrophage expression characteristics. In this cell model, TSP-1 protein expression is significantly increased and L-TGF-β is secreted; in the absence of factors that interfere with L-TGF-β activation, the plasmin secreted by the cells will recognize and cleave the TSP-1 / L-TGF-β complex bound to the cell surface CD36 receptor, thereby producing an increased amount of active TGF-β.

[0290] The assay was performed as follows: THP-1 cells were seeded in 96-well plates and treated with 300 nM PMA to differentiate and adhere to the culture medium, forming macrophage-like cells. 100 μM peptide was added (an equal volume of peptide solvent, PBS, was used in the negative control group, and an equal concentration of peptide SEQ ID NO: 50 was used in the positive control group). After 24 hours of treatment, the cells were inoculated with a 293 reporter cell line, the 293-TGFβRes cell line (novoprotein, Catalog No. XCC03-1), used to detect TGF-β activity. Lysate containing a fluorescent substrate (Vazyme, Catalog No. DD1201-01) was added, and fluorescence intensity was measured using a multi-function microplate reader (TECAN, Model Infinite 200Pro). The inhibitory effect of the peptide on TGF-β activation was assessed by calculating the ratio of fluorescence signal between the peptide-treated group and the negative control group. The test results are shown in Table 3. In this cell model, the polypeptide has different degrees of inhibitory effect on TGF-β activity, indicating that the polypeptide of the present invention can inhibit TSP-1-dependent TGF-β activation.

[0291] Table 3 Inhibitory effect of peptides on TSP-1-dependent TGFβ activation in cell models

[0292] Example 4: Inhibition of active TGFβ content by polypeptides in a fibrosis mouse disease model

[0293] This example evaluates the inhibitory effect of a peptide on active TGF-β levels during the progression of fibrotic diseases. This example utilizes a bleomycin (BLM)-induced mouse model of pulmonary fibrosis. The BLM animal model boasts high similarity to the underlying pathological features, ease of operation, and excellent reproducibility. It is the most representative model of fibrosis mechanisms and is a recognized and recommended animal model for drug efficacy studies.

[0294] In this embodiment, 6-8 week old 18-20g male C57 / 6J mice were used as research subjects. Experimental animals were randomly divided into groups of 8 in each group. The experimental endpoint (Day 8) detected the expression of profibrotic factors in the model. The modeling and administration methods in this embodiment were all intratracheally administered using an aerosol intratracheal dosing device (Yuyan, model YAN30012). The administration operation was as follows (An Official American Thoracic Society Workshop Report: Use of Animal Models for the Preclinical Assessment of Potential Therapies for Pulmonary Fibrosis, Am J Respir Cell Mol Biol Vol 56, Iss 5, pp 667–679, May 2017): The experimental animals were anesthetized by atomization of isoflurane (Ruiwode, batch number 22041701) using a small animal anesthesia machine (Ruiwode, model R500IE). An animal in an appropriately anesthetized state was placed in a supine position with its head upward and its body tilted, suspended by its incisors on an operating platform (tilted approximately 60°). The operator, holding a tissue forceps in their right hand, gently grasped the tongue and pushed it outward and upward to open the animal's mandible. With their left hand, they held a laryngoscope (Yuyan, model SR310-RW) and slid the blade of the laryngoscope along the middle of the tongue into the oral cavity to the anterior part of the glottis, gently pressing upward against the tongue and its base. The needle of the applicator was then inserted through the oral cavity, along the laryngoscope, into the animal's glottis, and 50 μL of the liquid in the applicator was rapidly injected. On the day of modeling (Day 1), the experimental animals were randomly divided into groups and administered 0.7 USP / kg bleomycin solution (Hanhui Pharmaceutical, batch number 20080511) through the trachea for modeling; on Day 7, each polypeptide treatment group was given a single intratracheal administration of 50 μL polypeptide drug (3 mg / kg dose), and the model group was given an equal volume of solvent (normal saline, Kelun, batch number L221062501) as a control; 24 hours after administration, the animals were dissected to obtain lung tissue, and the total protein of the mouse lung tissue was extracted after liquid nitrogen grinding. The TGF-β ELISA detection kit (BOSTER, product number EK0515) was used to determine the active TGF-β content in the lung tissue of each group of mice, and the ratio of the active TGF-β content in the polypeptide treatment group to the model group was calculated to evaluate the effect of the polypeptide on the active TGF-β content. The results are shown in Table 4. In the fibrosis disease model, the active TGF-β content in the polypeptide administration group was reduced to varying degrees compared with the model group, indicating that the polypeptide of the present invention has an inhibitory effect on the increase in active TGF-β content induced by lung injury during the progression of fibrosis.

[0295] Table 4 Inhibition of active TGF-β content in fibrosis mouse model by peptides

[0296] Example 5 Inhibition of TGFβ Downstream Signaling Pathway by Peptides in Fibrosis Mouse Disease Model

[0297] This example evaluates the inhibitory effect of the peptide on TGF-β downstream signaling during the progression of fibrotic diseases. TGF-β intracellular signaling depends on the phosphorylation of Smad family proteins, with increased Smad2 / 3 phosphorylation being a key marker for TGF-β signaling.

[0298] In this example, 6-8 week old male C57 / 6J mice weighing 18-20 g were used as the research subjects. The experimental animals were randomly divided into 8 groups. The modeling and drug administration methods in this example were both performed using an aerosol intratracheal dosing device for intratracheal administration, and the operation method was the same as described in Example 4. On the day of modeling (Day 1), the experimental animals were randomly divided into groups and administered 0.7 USP / kg bleomycin solution (Hanhui Pharmaceutical, batch number 20080511) through the trachea for modeling; on Day 7, each peptide-treated group was given a single 3 mg / kg dose of peptide drug through the trachea, and the BLM model group was given an equal volume of solvent (normal saline) as a control; 24 hours after administration, the animals were dissected and the lung tissue was frozen in liquid nitrogen for later use; after liquid nitrogen grinding, the total protein of the mouse lung tissue was extracted, and the P-Smad2 / 3 ELISA detection kit (CST, product number 12001C) was used to determine the P-Smad2 / 3 content in the total tissue. The ratio of P-Smad2 / 3 content in the peptide-treated group to the model group was calculated to evaluate the effect of the peptide on the TGF-β signaling pathway transduction. The results are shown in Table 5. In the fibrosis disease model, the P-Smad2 / 3 protein content in the polypeptide administration group was reduced to varying degrees compared with the model group, indicating that the polypeptide of the present invention has varying degrees of inhibition on the transduction of the TGF-β signaling pathway during the progression of fibrosis.

[0299] Table 5 Inhibition of TGF-β signaling pathway transduction by peptides in fibrosis mouse model

[0300] Example 6: Inhibition of collagen deposition by polypeptides in a fibrosis mouse disease model

[0301] This example is to evaluate the inhibitory effect of polypeptides on collagen deposition in fibrotic tissue. In this example, 6-8 week old 18-20g male C57 / 6J mice were used as research subjects, and the experimental animals were randomly divided into groups of 8 in each group. On the day of modeling (Day 1), a single 0.7USP / kg dose of bleomycin solution was administered through the trachea to establish the model; Day 8 began to administer treatment, and each polypeptide administration treatment group was given a 3mg / kg dose of polypeptide drug through the trachea, with a frequency of twice a week, for a total of 4 times. The BLM model group was given an equal volume of solvent (normal saline) as a control, and the operation method was the same as described in Example 4. Day 21 was used as the experimental endpoint to detect the degree of fibrosis in the model.

[0302] On Day 21, lung tissue was dissected from the animals and ground with liquid nitrogen before acid hydrolysis to extract hydroxyproline (HYP) content. HYP is a unique amino acid found in collagen, and its content is a key indicator of collagen metabolism and fibrosis. For every 100 mg of tissue, 1 mL of 6N HCl was added and mixed thoroughly. The tissue was hydrolyzed at 121°C for 6 hours. The supernatant of the hydrolyzate or a HYP standard (Sigma, Catalog No. V900395-25G) solution with varying concentrations was centrifuged and used for HYP content determination. In a 96-well plate, 10 μL of the hydrolyzate supernatant was added to each well and dried in a 65°C oven; 100 μL of 1.4% chloramine T (Sigma, Cat. No. 23270-50G) oxidizing solution was added to each well and incubated at room temperature for 15 minutes; 100 μL of Ehrlich colorimetric solution (Solar Bio, Cat. No. G1290) was added to each well and incubated at 65°C for 30 minutes; the absorbance value (OD560) of each sample well at 560 nm was detected using a multifunctional microplate reader (TECAN, Model Infinite 200Pro). The total HYP content in the lung tissue of each test group was calculated, and the reduction ratio of HYP content in the polypeptide administration group relative to the model group was obtained to evaluate the inhibitory effect of the polypeptide molecule on collagen deposition in the pathological site of fibrosis. The results are shown in Table 6. In the fibrosis disease model, the HYP content in the polypeptide administration group was reduced to varying degrees compared with the model group, indicating that the polypeptide of the present invention has varying degrees of inhibition on collagen deposition in fibrotic tissue.

[0303] Table 6 Inhibition of collagen deposition in fibrotic tissue by peptides

[0304] Example 7 Inhibitory Effects of Peptides on Inflammatory and Fibrotic Pathology in a Mouse Model of Fibrosis

[0305] This example is to evaluate the inhibitory effect of polypeptides on fibrotic pathology. Male C57 / 6J mice aged 6-8 weeks and weighing 18-20g were used as research subjects, and the experimental animals were randomly divided into groups of 6 in each group. On the day of modeling (Day 1), the BLM model group and each treatment group were given a single 0.7USP / kg dose of BLM solution through the trachea for modeling, and the negative control group was given an equal volume of normal saline. Dosing began on Day 8, and each polypeptide-administered treatment group was given a 3mg / kg dose of polypeptide drug through the trachea, and the positive control group was given an equal dose of polypeptide SEQ ID NO: 50. The dosing frequency was twice a week, for a total of 4 times; the BLM model group and the negative control group were given an equal volume of solvent (normal saline) as a control. The operating method is the same as described in Example 4.

[0306] On Day 21, lung tissue was dissected and washed with saline to remove blood. After drying with gauze, the dissected lung tissue was subjected to histopathological analysis. Lung tissue was perfused intratracheally with 0.5 mL of paraformaldehyde and placed in 4 mL of paraformaldehyde. A small amount of gauze was added to completely immerse the lung tissue in the fixative and fixed for at least 24 hours. The largest coronal section was trimmed from one lobe on the left and all four lobes on the right lung. The tissue was placed in an embedding cassette, dehydrated overnight, and then embedded in paraffin to prepare 3 μm sections. An automatic slide stainer (Leica, model ST5010) was used for hematoxylin (Heter, batch number 201905)-eosin (Cologne, batch number 2018070301) (Hematoxylin-Eosin, HE) staining and MASSON'S trichrome staining (Lazurite Blue, Cologne, batch number MKCH8129; Aniline Blue, Source Leaf, batch number H70J11S115483; Ponceau Red, Sigma, batch number SHBM2047). After sealing, the slides were scanned using a digital panoramic scanner (Zhiyue, model WS-10). ≤10 fields of view were captured for each lung lobe at 20x magnification. The severity of inflammation and fibrosis were scored by HE staining and MASSON'S trichrome staining, respectively, to evaluate the inhibitory effect of the peptide on inflammatory infiltration and fibrosis deposition in fibrotic pathological sites. The results are shown in Figures 1 and 2.

[0307] Figure 1 shows a pathological section stained with HE (20x magnification). The lung tissue in the negative control group showed thin-walled, vacuolar structures in the alveolar cavity. The BLM model group showed a large infiltration of inflammatory cells and fibroblasts in the interstitium and alveoli, accompanied by significant thickening of the alveolar interstitium and compression and deformation of the alveolar structure. All peptide-administered groups showed varying degrees of reduction in inflammatory cells and a reduction in the area of ​​thickened alveolar septa.

[0308] Figure 2 shows a MASSON'S-stained pathological section (20x magnification). In the negative control group, the collagen layer of the bronchial wall of mouse lung tissue was thin, and a small amount of strip-shaped, elongated collagen fibers were visible between the alveoli. In the BLM model group, the alveolar septa widened, with abundant collagen fiber deposition, and fibrotic masses formed in some areas. In each peptide-administered group, only a small amount of alveolar septa widened, and the area of ​​collagen fiber deposition was reduced to varying degrees compared to the BLM model group.

[0309] The results of the inflammatory score and fibrosis score are shown in Table 7. Compared with the negative control group, the inflammatory score and fibrosis score of the BLM model group were significantly increased, and the inflammatory score and fibrosis score of each peptide administration group decreased to varying degrees compared with the model group.

[0310] Table 7 Inhibition of inflammatory and fibrotic pathology by peptides in mouse fibrosis model

[0311] Example 8 Inhibition of Collagen Deposition by Peptides in a Rat Fibrosis Disease Model

[0312] This example is to evaluate the inhibitory effect of polypeptides on collagen deposition in pathological sites of pulmonary fibrosis. Male SD rats aged 6-8 weeks and weighing 200-220g were used as research subjects, and the experiment was randomly divided into groups with 8 rats in each group. On the day of modeling (Day 1), the model group and each treatment group were given a single intratracheal administration of 1.8USP / kg bleomycin solution for modeling; each polypeptide-administered treatment group began to be given a 2mg / kg dose of polypeptide through the trachea from Day 1, with a frequency of 2 times a week, for a total of 4 times. The model group was given an equal volume of solvent (normal saline) as a control, and the volume was 100μL. The operation method was the same as described in Example 4. Day 15 was used as the experimental endpoint to detect the degree of fibrosis in the model.

[0313] On Day 15, lung tissue was dissected and ground with liquid nitrogen. Total hydroxyproline (HYP) was extracted from rat lung tissue using acid hydrolysis. HYP content was determined using the chloramine T oxidation colorimetric method. 1 mL of 6N HCl was added to every 100 mg of tissue, mixed thoroughly, and hydrolyzed at 121°C for 6 h. The supernatant was centrifuged and used for HYP content determination. In a 96-well plate, 10 μL of hydrolysate supernatant or HYP standard (Sigma, Catalog No. V900395-25G) solutions of varying concentrations were added to each well and dried in a 65°C oven. 100 μL of 1.4% chloramine T (Sigma, Catalog No. 23270-50G) oxidizing solution was added to each well and incubated at room temperature for 15 minutes. 100 μL of Ehrlich colorimetric solution (Solarbo, Catalog No. G1290) was added to each well and incubated at 65°C for 30 minutes. The absorbance at 560 nm (OD560) of each sample well was measured using a multifunctional microplate reader (TECAN, Model Infinite 200Pro). The total HYP content in lung tissue of each experimental group was calculated, and the percentage of HYP reduction in the peptide-treated group relative to the model group was calculated to assess the inhibitory effect of the peptide on collagen deposition in fibrotic pathological sites. The results are shown in Table 8. In the pulmonary fibrosis disease model, the HYP content in the polypeptide-administered groups was reduced to varying degrees compared with the model group, indicating that the polypeptide of the present invention inhibited collagen deposition in the pathological tissues of pulmonary fibrosis to varying degrees.

[0314] Table 8: Inhibition of collagen deposition in fibrosis models by peptides

[0315] Example 9: Inhibitory Effect of Peptides on Fibrosis Pathology in Fibrosis Rat Disease Model

[0316] The present embodiment is to assess the inhibitory effect of polypeptide administration on the expression level of pro-fibrotic factors in fibrotic pathological sites. Male SD rats aged 6-8 weeks and 200-220g were used as research subjects, and the experiment was randomly divided into groups of 6 in each group. On the day of modeling (Day 1), the BLM model group and each treatment group were given a single 1.8USP / kg dose (100 μL) bleomycin solution in the trachea for modeling, and the negative control group was given an equal volume of normal saline. Each polypeptide administration treatment group started to be given 2mg / kg dosage polypeptide in the trachea from Day 1, with a frequency of administration 2 times a week for a total of 4 times. The BLM model group and the negative control group were given an equal volume of solvent (normal saline) as a control, and the volume was 100 μL. The operating method is described in Example 4.

[0317] On Day 15, lung tissue was dissected and removed, washed with saline to remove blood stains, and the lung weight was weighed after being dipped in gauze to dry. The lung coefficient was calculated (lung coefficient = lung wet weight / body weight * 1000, the lung coefficient reflects the general conditions of the lung tissue, such as edema, inflammation and fibrosis). The dissected lung tissue was subjected to histopathological analysis. The lung tissue was perfused with 5 mL of paraformaldehyde through the trachea and placed in 40 mL of paraformaldehyde. A small amount of gauze was added to completely immerse the lung tissue in the fixative and fixed for more than 24 hours. The largest coronal plane was trimmed from the left lobe and the four lobes of the right lung. The left and right lungs were placed in two embedding boxes respectively, and 3 μm sections were made after paraffin embedding. MASSON'S staining was performed according to the method described in Example 7. After digital scanning, ≤10 fields of view were intercepted for each lobe at 10 times the field of view, and the fibrosis severity score was performed to evaluate the inhibitory effect of the polypeptide on fibrosis deposition in pathological sites of fibrosis.

[0318] The results are shown in Figure 3 (magnification 10x). In the negative control group, the collagen layer of the bronchial wall of the rat lung tissue was thin, and a small amount of strip-shaped, slender collagen fibers were distributed between the alveoli. In the BLM model group, the alveolar septa of the lung tissue were widened, and a large amount of collagen fibers were deposited, with fibrotic masses forming in some areas. In each peptide-administered group, a small amount of alveolar septa widened, and the area of ​​collagen fiber deposition was reduced to varying degrees compared with the BLM model group.

[0319] The fibrosis score results are shown in Table 9. Compared with the negative control group, the lung coefficient and fibrosis severity score of the BLM model group increased significantly; the lung coefficient and fibrosis score of each peptide administration group decreased to varying degrees compared with the model group.

[0320] Table 9 Inhibition of fibrosis pathology by peptides in rat fibrosis model

[0321] Example 10: Inhibitory test of the ability of polypeptides to inhibit tumor cell migration

[0322] TGF-β plays a dual role in the tumor microenvironment (TME). In the early stages of tumor development, the TGF-β signaling pathway induces apoptosis and inhibits tumor cell proliferation. In the late stages of tumor development, TGF-β acts as an immunosuppressive cytokine, suppressing the immune response and promoting tumor progression by regulating tumor cell motility and metastasis.

[0323] This example is to evaluate the effect of polypeptides on the migration ability of different tumor cells. The experiment used a transwell nested chamber (membrane pore size of 8 μm), inoculated THP-1 cells in the lower chamber and differentiated and adhered to the wall with 300nM PMA (Sigma, Product No. P1585), and then added with 100μM polypeptide (negative control group added an equal volume of solvent) for 48 hours. The upper chamber was inoculated with tumor cells, human lung adenocarcinoma cell A549 cells (American Type Culture Collection, ATCC, CCL-185) and co-cultured for 24 hours, or inoculated with human liver cancer cell HepG2 / C3A cells (ATCC, CRL-10741) and co-cultured for 24 hours, or inoculated with human breast cancer cell MCF7 cells (ATCC, HTB-22) and co-cultured for 48 hours. Afterwards, the upper chamber was removed and fixed with 70% ethanol for 10 minutes, followed by washing. The cells were then stained with Giemsa stain (Biyuntian, Cat. No. C0131) for 45 minutes. After rinsing 2-3 times with double-distilled water, unmigrated cells in the upper layer were removed with a cotton swab. After the chamber dried, it was observed under a microscope. Five randomly selected fields of view were counted at 100x magnification. The results, as shown in Table 10, showed that the peptide significantly inhibited the migration of human lung adenocarcinoma cells, human liver cancer cells, and human breast cancer cells.

[0324] Table 10 Inhibitory effect of peptides on tumor cell migration

[0325] Example 11 Inhibition of ECM-related gene expression by polypeptides in a liver fibrosis cell model

[0326] This example evaluates the inhibitory effect of the polypeptide on the expression of extracellular matrix (ECM)-related genes in a liver fibrosis cell model. The experiment used human hepatic stellate cells LX-2. Hepatic stellate cells are the main source of myofibroblasts in the liver. After activation, they transform into myofibroblasts, secrete ECM in the liver, and directly participate in the formation of liver fibrosis. Brief stimulation of LX-2 cells by TGF-β1 can initiate positive feedback regulation of TSP-1-TGF-β1 activation events, continuously promoting ECM formation and other related myofibroblast characteristics.

[0327] The experimental procedure is as follows. After stimulating LX-2 cells (Procell, CL-0560) with 20 ng / mL TGF-β1 (GenScript, Cat. No. Z03411) for 24 hours, 100 μM peptide was added and cultured for 48 hours. An equal volume of solvent was added to the model group, and LX-2 cells without any treatment served as a blank control. Afterwards, LX-2 cells were collected and total RNA was extracted for RT-qPCR to detect the expression of ECM-related genes, fibronectin EDA-Fibronectin (FN) and collagen I (COL1). The experimental test results are shown in Table 11. The peptide can reduce the expression of FN and COL1 genes in LX-2 and inhibit ECM formation.

[0328] Table 11 Inhibitory effect of peptides on ECM-related gene expression in liver fibrosis cell model

[0329] Sequence Listing Overview

[0330] The following table provides information about the peptides described by each SEQ ID NO in the sequence listing, where Ac represents an acetyl group and NH2 represents an amino group.

Claims

1. A peptide having an amino acid sequence of formula (I) or a pharmaceutically acceptable salt, solvate or prodrug thereof, R 1 -X 1 -X 2 -X 3 -X 4 -X 5 -X 6 -X 7 -R 2 (I), in, R 1 is acetyl or does not exist; R 2 It is amino or does not exist; X 1 Not present, or selected from T and S; X 2 Selected from natural amino acids or non-natural amino acids, preferably, selected from polar side chain amino acids, such as uncharged polar side chain amino acids S, C, G, N, Q, T, Y or negatively charged polar side chain amino acids D and E, more preferably selected from amino acids N, C, Q, S, T, E and D; more preferably selected from amino acids Q, C, N, E or S; X 3 It is D; X 4 is selected from amino acids with small side chains, such as A, C, G, P, S, T and V, more preferably selected from amino acids A, P and S, and more preferably selected from amino acids A and P; X 5 It is E; X 6 It is D; X 7 does not exist, or is selected from the amino acid sequence Z 1 Z 2 Z 3 SZ 4 LQ is a sequence of 1, 2, 3, 4, 5, 6 or 7 amino acids from the amino terminus to the carboxyl terminus. where Z 1 is the amino acid N, Q, P, A, L, V, M, or I, preferably N, P, or L, most preferably N or P; Z 2 is an amino acid T, S, V, C, A, K, or R, preferably T, C or K, more preferably, when Z 2 When it is amino acid C or K, Z 2 With X 2 or X 3 The amino acid side chains form covalent bonds, Z 3 is amino acid A, V, L, or I, preferably A or V, Z 4 is amino acid F, Y, H, or W, preferably F or H; Optionally, when X 4 When A, at X 3 and X 4 0-1 small side chain amino acids, such as P or G, are inserted between, preferably, P is inserted; Optionally, when X 7 When the peptide is composed of 1-5 amino acids, the peptide is 7 The amino acid residues may be followed by 0-2 additional amino acid residues, for example, 1-2 amino acid residues selected from the group consisting of Q, E, D, and N, or 1 amino acid residue selected from the group consisting of C or K.

2. The peptide according to claim 1 or a pharmaceutically acceptable salt, solvate or prodrug thereof, wherein X 1 Not present, or selected from T and S; X 2 selected from the group consisting of amino acids Q, N, C, S, T, E and D; X 3 It is D; X 4 Selected from A, C, G, P, S, T and V; X 5 It is E; X 6 It is D; X 7 does not exist, or is selected from the amino acid sequence Z 1 Z 2 Z 3 SZ 4 LQ is a sequence of 1, 2, 3, 4, 5, 6 or 7 amino acids from the amino terminus to the carboxyl terminus, wherein Z 1 is N, Q, P, A, L, V, M, or I; Z 2 is T, K or C; Z 3 is A or V; Z 4 It is F, Y, H, or W.

3. The peptide according to any one of claims 1 to 2, or a pharmaceutically acceptable salt, solvate or prodrug thereof, wherein: X 1 Selected from T and S; X 2 Selected from Q, N, E, S or C; X 3 It is D; X 4 Selected from A and P; X 5 It is E; X 6 It’s D.

4. The peptide according to any one of claims 1 to 3, or a pharmaceutically acceptable salt, solvate or prodrug thereof, wherein The peptide comprises an amino acid sequence selected from the group consisting of: -DAED; -DPED; -DAEDN; -DPEDN; or -DAEDP.

5. The peptide according to any one of claims 1 to 4, or a pharmaceutically acceptable salt, solvate or prodrug thereof, wherein X 7 from Z 1 、Z 1 Z 2 、Z 1 Z 2 Z 3 、Z 1 Z 2 Z 3 S, Z 1 Z 2 Z 3 SZ 4 、Z 1 Z 2 Z 3 SZ 4 L or Z 1 Z 2 Z 3 SZ 4 composed of LQ, preferably, X 7 from Z 1 Z 2 Z 3 、Z 1 Z 2 Z 3 S or Z 1 Z 2 Z 3 SZ 4 composed. And among them Z 1 is N, P or L; Z 2 is T, K or C; Z 3 is A or V; Z 4 It is F, Y, H, or W.

6. The peptide according to claim 5 or a pharmaceutically acceptable salt, solvate or prodrug thereof, wherein Z 1 is N or P; Z 2 is T, K or C; Z 3 A or V, preferably A; Z 4 It is F or H.

7. The peptide according to any one of claims 1 to 6, or a pharmaceutically acceptable salt, solvate or prodrug thereof, wherein: X 7 It is an amino acid sequence selected from the group consisting of: N; P; L; NTA; NTV; PTA; NTAS; PTAS; NTVSF; NTASF; NTASH; NTVSFQ.

8. The peptide according to any one of claims 1 to 7, or a pharmaceutically acceptable salt, solvate or prodrug thereof, wherein X 1 It’s T.

9. The peptide according to any one of claims 1 to 8, or a pharmaceutically acceptable salt, solvate or prodrug thereof, wherein: X 4 It’s A.

10. The peptide according to any one of claims 1 to 8, or a pharmaceutically acceptable salt, solvate or prodrug thereof, wherein X 4 It’s P.

11. The peptide according to any one of claims 1 to 10, or a pharmaceutically acceptable salt, solvate or prodrug thereof, wherein: Z 1 It's N.

12. The peptide according to any one of claims 1 to 10, or a pharmaceutically acceptable salt, solvate or prodrug thereof, wherein: Z 1 It’s P.

13. The peptide according to any one of claims 1 to 12, or a pharmaceutically acceptable salt, solvate or prodrug thereof, wherein: Z 3 It’s A.

14. The peptide according to any one of claims 1 to 13, or a pharmaceutically acceptable salt, solvate or prodrug thereof, wherein: X 2 It's Q.

15. The peptide according to any one of claims 1 to 14, or a pharmaceutically acceptable salt, solvate or prodrug thereof, wherein Z 2 It’s T.

16. The peptide according to any one of claims 1 to 13, or a pharmaceutically acceptable salt, solvate or prodrug thereof, wherein: X 2 It's C, Z 2 It's C, X 2 and Z 2 The amino acid side chains are covalently linked by disulfide bonds.

17. The peptide according to any one of claims 1 to 13, or a pharmaceutically acceptable salt, solvate or prodrug thereof, wherein: X 3 It is D, Z 2 It's K, X 3 and Z 2 The amino acid side chains of the α-amino acids are covalently linked to form lactam bonds.

18. The peptide according to any one of claims 1 to 17, or a pharmaceutically acceptable salt, solvate or prodrug thereof, wherein: R 1 It is acetyl.

19. The peptide according to any one of claims 1 to 18, or a pharmaceutically acceptable salt, solvate or prodrug thereof, wherein: R 2 It is an amino group.

20. The peptide according to any one of claims 1 to 19, or a pharmaceutically acceptable salt, solvate or prodrug thereof, wherein: The peptide consists of 5 to 13 amino acids, preferably 7, 8, 9, 10 or 11 amino acids.

21. The peptide according to any one of claims 1 to 20, or a pharmaceutically acceptable salt, solvate or prodrug thereof, wherein: The peptide is a linear peptide or a cyclic peptide.

22. The peptide according to any one of claims 1 to 21 or a pharmaceutically acceptable salt, solvate or prodrug thereof, wherein: The peptide is a cyclic peptide, preferably, the cyclic peptide is a head-to-tail ring, a DK side chain ring, or a disulfide bond ring, Wherein, the cyclization region of the peptide comprises a core motif (DXED), Preferably, the peptide is coupled to form a ring via the side chains of the amino acids at positions 3 and 8, or via the side chains of the amino acids at positions 2 and 8, or Preferably, wherein X 3 It's D, Z 2 is K, and the peptide is 3 With Z 2 cyclization of the lactam bond formed between the side chains of Preferably, wherein X 2 It's C, Z 2 is C, and the peptide is connected by X 2 With Z 2 The disulfide bonds formed between the side chains of More preferably, when Z 2 When it is amino acid C or K, X 7 By Z 1 Z 2 Composition, and X 1 T or S.

23. The peptide according to any one of claims 1 to 22, or a pharmaceutically acceptable salt, solvate or prodrug thereof, wherein: The peptide is chemically modified, for example, PEG-modified, lipid-modified, substituted with D-amino acids, or substituted with unnatural amino acids.

24. A peptide selected from any one of SEQ ID NOs: 1 to 49, or a pharmaceutically acceptable salt, solvate or prodrug thereof, or a peptide that differs from said peptide in amino acid sequence by one or two amino acid substitutions, deletions or additions, preferably, said substitutions, deletions and additions occur at amino acid residues outside the core motif (DXED), for example, conservative amino acid substitutions.

25. A peptide selected from any one of SEQ ID NOs: 6, 22, 25, 26, 33, 40, 43, 44 and 46, or a pharmaceutically acceptable salt, solvate or prodrug thereof.

26. A method for preparing the peptide according to any one of claims 1 to 25 or a pharmaceutically acceptable salt, solvate or prodrug thereof, preferably, the method comprises synthesizing the peptide by solid phase synthesis.

27. A pharmaceutical composition comprising the peptide according to any one of claims 1 to 25 or a pharmaceutically acceptable salt, solvate or prodrug thereof, and a pharmaceutically acceptable carrier.

28. A method for preventing or treating a TGF-β-related disease, comprising administering to a subject in need thereof a preventively or therapeutically effective amount of a peptide according to any one of claims 1 to 25 or a pharmaceutically acceptable salt, solvate or prodrug thereof, or a pharmaceutical composition according to claim 27, Optionally, the method further comprises administering to the subject an effective amount of a second therapeutic agent; Preferably, the disease has macrophage infiltration at the lesion site, or is associated with tissue damage, inflammation or fibrosis.

29. The method of claim 28, wherein the TGF-β related disease is a fibrotic disorder, preferably selected from: pulmonary fibrosis (eg, IPF), liver fibrosis, kidney fibrosis, myelofibrosis, myocardial fibrosis, and / or skin fibrosis.

30. The method according to claim 28, wherein the TGF-β related disease is a solid tumor, preferably selected from the group consisting of lung cancer, liver cancer, breast cancer, uterine cancer, prostate cancer, pancreatic cancer, colon cancer, skin cancer, central nervous system cancer, fibromyoma, fibroma, fibroadenoma and fibrosarcoma.

31. Use of a peptide according to any one of claims 1 to 25, or a pharmaceutically acceptable salt, solvate or prodrug thereof, for in vitro or in vivo: - Blocking the binding of CD36 to TSP-1; - Inhibits TSP-1-dependent TGF-β1 activation; - Reduce the amount of active TGF-β1 in fibrotic tissue; -Inhibits TGF-β1-mediated downstream signaling; - Reduce collagen deposition in fibrotic tissue; - Inhibit inflammatory and / or fibrotic lesions mediated by TGF-β1; - Inhibits the expression of extracellular matrix-related genes stimulated by TGF-β1; -Inhibit tumor cell migration; -Prevention or treatment of TGF-β1-related diseases, especially fibrosis or cancer; Or used for preparing medicines used in the above purposes.

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