TGF-beta receptor agonist polypeptides and uses thereof

Abstract

The invention belongs to the field of polypeptide medicines, and particularly relates to a TGF-beta receptor agonist polypeptide and application thereof. The amino acid sequence of the polypeptide is as shown in SEQ ID No. 1. The polypeptide disclosed by the invention has a remarkable agonistic effect on a TGF-beta receptor, and can be applied to preparation of medicines for preventing and / or treating diseases related to TGF-beta, in particular to prevention of alopecia and / or promotion of hair growth.

Description

Technical Field

[0001] The present invention belongs to the field of polypeptide medicine, and specifically relates to a TGF-β receptor agonist polypeptide and its application. Background Art

[0002] Transforming growth factor beta (TGF-β) is a member of the TGF-β superfamily. TGF-β includes three subtypes, TGF-β1, TGF-β2 and TGF-β3. TGF-β is a multifunctional protein. A large number of studies have shown that TGF-β has different functions in different cell types, and this function depends on the specific environment. Normally, cell survival, metabolism, growth, proliferation, differentiation, adhesion, migration and death are all regulated by TGF-β. Normal TGF-β signaling is essential for the normal function and homeostasis of a healthy body, while abnormal TGF-β signaling may lead to various diseases.

[0003] TGF-β has a high affinity receptor (TGF-βR) on the cell surface, which is divided into three subtypes: TGF-βRI, TGF-βRII, and TGF-βRIII. TGF-β forms a complex by binding to TGF-βRII with serine / threonine kinase activity on the cell membrane surface, and then forms a complex with TGF-βRI, which then transmits signals and activates the corresponding signaling pathways, thereby regulating cell proliferation, differentiation, and apoptosis.

[0004] Today, TGF-β remains a challenging therapeutic target due to its diverse functions, variable biological behaviors, and toxicity in organs such as the heart. Despite the many challenges, in recent years, the emergence of concepts such as small molecule inhibitors, fusion molecules, monoclonal antibodies, "TGF-β traps", vaccines, and antisense oligonucleotide molecules has made significant progress in the development of therapeutics targeting the TGF-β pathway.

[0005] Prior art CN104902916A discloses a mutant polypeptide of the TGF-β molecule, whose primary sequence is highly homologous to the sequence of human TGF-β. The mutant polypeptide loses the ability to interact with TGF-βRI (also known as ALK5) but retains the interaction with the receptors TGF-βRII and TGF-βRIII (see paragraphs [0015-0017] of the specification); existing TGF-βRI inhibitors, including SM16 and GW788388, have been shown to have anti-fibrotic effects; in addition, including Galunisertib, PF-06952229, LY2109761, etc., specifically bind to TGF-βR1, prevent TGF-βR1-mediated signal transduction, and are thus used for anti-tumor treatment.

[0006] Qixuan Wang's team at the University of California, Riverside, determined how TGF-β controls the division and formation of cells (including stem cells) in hair follicles, or controls their own death and ultimately leads to the death of the entire hair follicle (A probabilistic Boolean model on hair follicle cell fate regulation by TGF-β, Biophys J. 2022 Jul 5; 121 (13): 2638-2652.). The results showed that transforming growth factor β (TGF-β) exhibits a dual role in regulating the fate of hair follicle cells, which can be anti-apoptotic or pro-apoptotic. By activating the TGF-β signaling pathway, it helps to activate hair follicle cells to produce new life, thereby activating hair follicle stem cells and stimulating hair growth. Summary of the invention

[0007] After in-depth research and creative work, the present applicant has discovered a group of polypeptides that can effectively activate TGF-β receptors, can be artificially intervened to activate hair follicle stem cells and stimulate hair growth, and the complete healing of wounds also requires the regeneration of hair follicles, and has the potential to be used as a therapeutic or auxiliary therapeutic drug. The following invention is provided:

[0008] One aspect of the present invention relates to a polypeptide or a pharmaceutically acceptable salt thereof, wherein the amino acid sequence of the polypeptide is as shown in SEQ ID No. 1:

[0009] ALX 3 TNYCFSSTX 12 TNCCVRPLYIDFEKDLGWKWIHEPKGYX 40 ANFCLGPCPX 50 GWSLX 55TQYSKVLALYX 66 X 67 X 68 NPX 71 SSAX 75 PCCVPX 81 X 82 LX 84 PLX 87 IX 89 YYX 92 GX 94 X 95 X 96 X 97 TX 99 QLSNMIVX 107 X 108 CX 110 CS

[0010] in,

[0011] X 3 indicates D, W or Y;

[0012] X 12 Indicates E or W;

[0013] X 40 Indicates E or Y;

[0014] X 50 Indicates P or S;

[0015] X 55 Indicates R, E or S;

[0016] X 66 Indicates K or H;

[0017] X 67 It means E, S or A;

[0018] X 68 Indicates H or R;

[0019] X 71 Indicates S, Q or T;

[0020] X 75 represents S, A, R or K;

[0021] X 81 Indicates S, E, R or K;

[0022] X 82 Indicates V, K, E or S;

[0023] X 84 means A, E or S;

[0024] X 87 Indicates T or E;

[0025] X 89 represents K, T or E;

[0026] X 92 Indicates K or V;

[0027] X 94 Indicates K, D, N, R or L;

[0028] X 95 Indicates K or T;

[0029] X 96 Indicates L or K;

[0030] X 97 Indicates Q, V or I;

[0031] X 99 Indicates K or E;

[0032] X 107 Indicates L, Q, K or E;

[0033] X 108 Indicates S or A;

[0034] X 110 Indicates T, R or V;

[0035] When X 66 Indicates H, X 67 Indicates A, X 68 When H is indicated, X 94 Not for R;

[0036] When X 66 Indicates H, X 67 Indicates A, X 68 Indicates H, X 71 When X represents S, 94 When it is not N.

[0037] In some examples, the amino acid sequence of the polypeptide is as shown in SEQ ID No. 2, SEQ ID No. 2:

[0038] ALX 3 TNYCFSSTX 12 TNCCVRPLYIDFEKDLGWKWIHEPKGYX 40 ANFCLGPCPX 50 GWSL X 55 TQYSKVLALYX 66 X 67 X 68 NPX 71 SSAX 75 PCCVPX81 X 82 LX 84 PLX 87 IX 89 YYX 92 GX 94 X 95 X 96 X 97 TKQLSNMIVX 107 X 108 CX 110 CS

[0039] in,

[0040] X 3 Indicates D or W;

[0041] X 12 Indicates E or W;

[0042] X 40 Indicates E or Y;

[0043] X 50 Indicates P or S;

[0044] X 55 Indicates R, E or S;

[0045] X 66 Indicates K or H;

[0046] X 67 It means E, S or A;

[0047] X 68 Indicates H or R;

[0048] X 71 Indicates S, Q or T;

[0049] X 75 represents S, A, R or K;

[0050] X 81 Indicates S, E, R or K;

[0051] X 82 Indicates V, K or S;

[0052] X 84 means A, E or S;

[0053] X 87 Indicates T or E;

[0054] X 89 represents K, T or E;

[0055] X 92 Indicates K or V;

[0056] X 94 Indicates K, D, N, R or L;

[0057] X 95 Indicates K or T;

[0058] X 96 Indicates L or K;

[0059] X 97 Indicates Q, V or I;

[0060] X 107 Indicates L, Q, K or E;

[0061] X 108 Indicates S or A;

[0062] X 110 Indicates T, R;

[0063] When X 66 Indicates H, X 67 Indicates A, X 68 When H is indicated, X 94 Not for R;

[0064] When X 66 Indicates H, X 67 Indicates A, X 68 Indicates H, X 71 When X represents S, 94 When it is not N.

[0065] In some examples, the amino acid sequence of the polypeptide is as shown in SEQ ID No. 3, SEQ ID No. 3:

[0066] ALX 3 TNYCFSSTX 12 TNCCVRPLYIDFEKDLGWKWIHEPKGYX 40 ANFCLGPCPX 50 GWSL X 55 TQYSKVLALYX 66 X 67 X 68 NPX 71 SSAX 75 PCCVPX 81 X 82 LX 84 PLX 87 IX 89 YYKG 94 KX 96 X 97 TKQ LSNMIVX 107 SCX110 CS

[0067] in,

[0068] X 3 Indicates D or W;

[0069] X 12 Indicates E or W;

[0070] X 40 Indicates E or Y;

[0071] X 50 Indicates P or S;

[0072] X 55 Indicates E or S;

[0073] X 66 Indicates K or H;

[0074] X 67 Indicates S or A;

[0075] X 68 Indicates H or R;

[0076] X 71 Indicates S or Q;

[0077] X 75 Indicates A, R or K;

[0078] X 81 Indicates S, E or K;

[0079] X 82 Indicates V or K;

[0080] X 84 Indicates A or E;

[0081] X 87 Indicates T or E;

[0082] X 89 Indicates K or T;

[0083] X 94 Indicates K, D, R or L;

[0084] X 96 Indicates L or K;

[0085] X 97 Indicates V or I;

[0086] X 107 Indicates Q, K or E;

[0087] X 110 Indicates T or R;

[0088] When X66 Indicates H, X 67 Indicates A, X 68 When H is indicated, X 94 Not for R;

[0089] When X 66 Indicates H, X 67 Indicates A, X 68 Indicates H, X 71 When X represents S, 94 When it is not N.

[0090] Preferably, in some examples, X 3 Indicates D.

[0091] Preferably, in some examples, X 12 Indicates E.

[0092] Preferably, in some examples, X 40 Indicates Y.

[0093] Preferably, in some examples, X 50 Indicates P.

[0094] Preferably, in some examples, X 55 Indicates E.

[0095] Preferably, in some examples, X 66 Indicates H.

[0096] Preferably, in some examples, X 67 Indicates A.

[0097] Preferably, in some examples, X 71 Indicates S.

[0098] Preferably, in some examples, X 75 Indicates K.

[0099] Preferably, in some examples, X 81 Indicates E.

[0100] Preferably, in some examples, X 82 Indicates K.

[0101] Preferably, in some examples, X 84 Indicates A.

[0102] Preferably, in some examples, X 87 Indicates T.

[0103] Preferably, in some examples, X 89 Indicates T.

[0104] Preferably, in some examples, X94 Indicates K or R.

[0105] Preferably, in some examples, X 96 Indicates K.

[0106] Preferably, in some examples, X 97 Indicates V.

[0107] Preferably, in some examples, X 107 Indicates E.

[0108] Preferably, in some examples, X 110 Indicates R.

[0109] In some examples, the amino acid sequence of the polypeptide is as shown in SEQ ID No.4, SEQ ID No.4:

[0110] ALDTNYCFSSTETNCCVRPLYIDFEKDLGWKWIHEPKGYYANFCLGPCPPGWSLETQY SKVLALYHAX 68 NPSSSAKPCCVPEKLAPLTITYYKGX 94 KKVTKQLSNMIVESCRCS

[0111] in,

[0112] X 68 Indicates H or R;

[0113] X 94 Indicates K or R.

[0114] In some examples, the amino acid sequence of the polypeptide is any one of SEQ ID No.5 to SEQ ID No.16.

[0115] SEQ ID No.5:

[0116] ALDTNYCFSSTETNCCVRPLYIDFEKDLGWKWIHEPKGYYANFCLGPCPSGWSLRTQY SKVLALYKEHNPSSSSASPCCVPRVLAPLTIKYYKGKKLQTKQLSNMIVLSCTCS,

[0117] SEQ ID No.6:

[0118] ALDTNYCFSSTETNCCVRPLYIDFEKDLGWKWIHEPKGYEANFCLGPCPPGWSLETQY SKVLALYKSHNPQSSAAPCCVPKVLAPLEIKYYKGKKLVTKQLSNMIVQSCTCS、

[0119] SEQ ID No.7:

[0120] ALDTNYCFSSTETNCCVRPLYIDFEKDLGWKWIHEPKGYYANFCLGPCPPGWSLSTQY SKVLALYHSHNPSSSARPCCVPSVLEPLTITYYKGDKKITKQLSNMIVKSCTCS、

[0121] SEQ ID No.8:

[0122] ALDTNYCFSSTETNCCVRPLYIDFEKDLGWKWIHEPKGYYANFCLGPCPPGWSLRTQY SKVLALYHAHNPTSSARPCCVPRKLAPLTIEYYVGNKKVTKQLSNMIVQSCTCS、

[0123] SEQ ID No.9:

[0124] ALDTNYCFSSTETNCCVRPLYIDFEKDLGWKWIHEPKGYYANFCLGPCPPGWSLETQY SKVLALYHAHNPSSSAKPCCVPEKLAPLTITYYKGKKKVTKQLSNMIVESCRCS、

[0125] SEQ ID No.10:

[0126] ALDTNYCFSSTETNCCVRPLYIDFEKDLGWKWIHEPKGYYANFCLGPCPPGWSLRTQY SKVLALYKSHNPQSSAAPCCVPRSLSPLTITYYVGNTKITKQLSNMIVQACTCS、

[0127] SEQ ID No.11:

[0128] ALWTNYCFSSTETNCCVRPLYIDFEKDLGWKWIHEPKGYYANFCLGPCPPGWSLETQY SKVLALYHAHNPSSSAKPCCVPEKLAPLTITYYKGKKKVTKQLSNMIVESCRCS、

[0129] SEQ ID No.12:

[0130] ALDTNYCFSSTWTNCCVRPLYIDFEKDLGWKWIHEPKGYYANFCLGPCPPGWSLETQY SKVLALYHAHNPSSSAKPCCVPEKLAPLTITYYKGKKKVTKQLSNMIVESCRCS、

[0131] SEQ ID No.13:

[0132] ALDTNYCFSSTETNCCVRPLYIDFEKDLGWKWIHEPKGYYANFCLGPCPPGWSLETQY SKVLALYHARNPSSSAKPCCVPEKLAPLTITYYKGKKKVTKQLSNMIVESCRCS、

[0133] SEQ ID No.14:

[0134] ALDTNYCFSSTETNCCVRPLYIDFEKDLGWKWIHEPKGYYANFCLGPCPPGWSLETQY SKVLALYHARNPSSSAKPCCVPEKLAPLTITYYKGLKKVTKQLSNMIVESCRCS、

[0135] SEQ ID No.15:

[0136] ALDTNYCFSSTETNCCVRPLYIDFEKDLGWKWIHEPKGYYANFCLGPCPPGWSLETQY SKVLALYHARNPSSSAKPCCVPEKLAPLTITYYKGRKKVTKQLSNMIVESCRCS。

[0137] Preferably, the amino acid sequence of the polypeptide is any one of SEQ ID No.6, SEQ ID No.7, SEQ ID No.9, SEQ ID No.11, SEQ ID No.13, SEQ ID No.14, SEQ ID No.15.

[0138] More preferably, the amino acid sequence of the polypeptide is any one of SEQ ID No.9, SEQ ID No.13, and SEQ ID No.15.

[0139] When used in the present invention, site-directed mutagenesis technology is used to identify amino acids that play a key role in the activity of the peptide of the present invention, and to determine the amino acids that play a key role in the activity of the polypeptide molecule.

[0140] Site-directed mutagenesis techniques, such as alanine scanning, introduce mutations into every residue of the polypeptide molecule, and test the biological activity of the resulting polypeptide molecule to determine the amino acid residues that play a key role in the activity of the polypeptide molecule; such as PCR site-directed mutagenesis techniques, by designing primers containing non-specific paired bases, and then introducing mutation sites into the product through PCR.

[0141] Through site-directed mutagenesis technology, it was determined that E at position 25 of the polypeptide of the present invention is an amino acid necessary for activity, further, "YIDFEKDLGW" at positions 20 to 30 are amino acids necessary for activity, and further, "TNCCVRPLYIDFEKDLGWKWIHEPKGY" at positions 13 to 39 are amino acids necessary for activity.

[0142] In another aspect, the present invention provides a recombinant polypeptide or a pharmaceutically acceptable salt thereof, wherein the recombinant polypeptide is obtained by coupling a cell penetrating peptide and the polypeptide described in SEQ ID No. 1 via a linker, wherein the coupling is covalently linked via an amide bond.

[0143] In some examples, the linker comprises, for example, at least 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 amino acid residues.

[0144] In some examples, the linker is selected from one of G, GG, GGG, S, SS, SSS, GS, GSGS, GSGSGS, GGS, GGSGGS, GGSGGSGG, GGGS, GGGSGGGSGGGS, GSS, GSSGSS, GSSGSSGSS, GGSS, GGSSGGSS, and GGSSGGSSGGSS.

[0145] Preferably, the linker is selected from GSGS.

[0146] In some examples, the cell penetrating peptide is selected from any one or more of TAT peptide of HIV-1 virus reverse transcription activator protein, D-Tat, transcription factor Antp peptide of Drosophila antennal homeotic protein, Pep-1 peptide, Pep-2 peptide, MPG peptide and RGD peptide.

[0147] In some examples, the cell penetrating peptide is selected from the group consisting of:

[0148] CPP sequence HIVTAT GRKKRRQRRRPQ cyclizableHIVTAT CGRKKRRQRRRPQC HA-TAT GDIMGEWGNEIFGAIAGFLGGRKKRRQRRRPQ Cell-penetrating peptides RQIKIWFQNRRMKWKK Cyclizable cell-penetrating peptide CRQIKIWFQNRRMKWKKC Transporter GWTLNSAGYLLGKINLKALAALAKKIL Cyclotransporter CGWTLNSAGYLLGKINLKALAALAKKILC Xentry LCLRPVG Cyclizable Xentry CLCLRPVGC R8 RRRRRRRR Cyclizable R8 CRRRRRRRRC

[0149] Preferably, in some examples, the cell penetrating peptide is selected from GRKKRRQRRRPQ.

[0150] In some examples, the recombinant polypeptide is obtained by coupling a cell-penetrating peptide and a polypeptide shown in SEQ ID No. 1 via a linker.

[0151] In some examples, the recombinant polypeptide is obtained by coupling a cell-penetrating peptide and a polypeptide shown in SEQ ID No. 2 via a linker.

[0152] In some examples, the recombinant polypeptide is obtained by coupling a cell-penetrating peptide and a polypeptide shown in SEQ ID No. 3 via a linker.

[0153] In some examples, the recombinant polypeptide is obtained by coupling a cell-penetrating peptide and a polypeptide shown in SEQ ID No. 4 via a linker.

[0154] In some examples, the recombinant polypeptide is obtained by coupling a cell-penetrating peptide and a polypeptide shown by SEQ ID No. 5 to SEQ ID No. 15 via a linker.

[0155] In some examples, the cell penetrating peptide is connected to the N-terminus or C-terminus of the above polypeptide via a linker.

[0156] In some examples, the cell penetrating peptide is located at the N-terminus of the recombinant polypeptide.

[0157] In some examples, the cell penetrating peptide is located at the C-terminus of the recombinant polypeptide.

[0158] In some examples, the amino acid sequence of the recombinant polypeptide is shown in SEQ ID No. 16, SEQ ID No. 16:

[0159] GRKKRRQRRRPQGSGSALX 3 TNYCFSSTX 12 TNCCVRPLYIDFEKDLGWKWIHEPKGYX 40 ANFCLGPCPX 50 GWSLX 55 TQYSKVLALYX 66 X 67 X 68 NPX 71 SSAX 75 PCCVPX 81 X 82 LX 84 PLX 87 IX89 YYX 92 GX 94 X 95 X 96 X 97 TX 99 QLSNMIVX 107 X 108 CX 110 CS

[0160] in,

[0161] X 3 indicates D, W or Y;

[0162] X 12 Indicates E or W;

[0163] X 40 Indicates E or Y;

[0164] X 50 Indicates P or S;

[0165] X 55 Indicates R, E or S;

[0166] X 66 Indicates K or H;

[0167] X 67 It means E, S or A;

[0168] X 68 Indicates H or R;

[0169] X 71 Indicates S, Q or T;

[0170] X 75 represents S, A, R or K;

[0171] X 81 Indicates S, E, R or K;

[0172] X 82 Indicates V, K, E or S;

[0173] X 84 means A, E or S;

[0174] X 87 Indicates T or E;

[0175] X 89 represents K, T or E;

[0176] X 92 Indicates K or V;

[0177] X 94 Indicates K, D, N, R or L;

[0178] X 95 Indicates K or T;

[0179] X 96 Indicates L or K;

[0180] X 97 Indicates Q, V or I;

[0181] X 99 Indicates K or E;

[0182] X 107 Indicates L, Q, K or E;

[0183] X 108 Indicates S or A;

[0184] X 110 Indicates T, R or V;

[0185] When X 66 Indicates H, X 67 Indicates A, X 68 When H is indicated, X 94 Not for R;

[0186] When X 66 Indicates H, X 67 Indicates A, X 68 Indicates H, X 71 When X represents S, 94 When it is not N.

[0187] In some examples, the amino acid sequence of the recombinant polypeptide is as shown in SEQ ID No. 17, SEQ ID No. 17:

[0188] GRKKRRQRRRPQGSGSALX 3 TNYCFSSTX 12 TNCCVRPLYIDFEKDLGWKWIHEPKGYX 40 ANFCLGPCPX 50 GWSLX 55 TQYSKVLALYX 66 X 67 X 68 NPX 71 SSAX 75 PCCVPX 81 X 82 LX 84 PLX 87 IX 89 YYX 92 GX 94 X 95 X96 X 97 TKQLSNMIVX 107 X 108 CX 110 CS

[0189] in,

[0190] X 3 Indicates D or W;

[0191] X 12 Indicates E or W;

[0192] X 40 Indicates E or Y;

[0193] X 50 Indicates P or S;

[0194] X 55 Indicates R, E or S;

[0195] X 66 Indicates K or H;

[0196] X 67 It means E, S or A;

[0197] X 68 Indicates H or R;

[0198] X 71 Indicates S, Q or T;

[0199] X 75 represents S, A, R or K;

[0200] X 81 Indicates S, E, R or K;

[0201] X 82 Indicates V, K or S;

[0202] X 84 means A, E or S;

[0203] X 87 Indicates T or E;

[0204] X 89 represents K, T or E;

[0205] X 92 Indicates K or V;

[0206] X 94 Indicates K, D, N, R or L;

[0207] X 95 Indicates K or T;

[0208] X 96Indicates L or K;

[0209] X 97 Indicates Q, V or I;

[0210] X 107 Indicates L, Q, K or E;

[0211] X 108 Indicates S or A;

[0212] X 110 Indicates T, R;

[0213] When X 66 Indicates H, X 67 Indicates A, X 68 When H is indicated, X 94 Not for R;

[0214] When X 66 Indicates H, X 67 Indicates A, X 68 Indicates H, X 71 When X represents S, 94 When it is not N.

[0215] In some examples, the amino acid sequence of the recombinant polypeptide is shown in SEQ ID No. 18, SEQ ID No. 18:

[0216] GRKKRRQRRRPQGSGSALX 3 TNYCFSSTX 12 TNCCVRPLYIDFEKDLGWKWIHEPKGYX 40 ANFCLGPCPX 50 GWSLX 55 TQYSKVLALYX 66 X 67 X 68 NPX 71 SSAX 75 PCCVPX 81 X 82 LX 84 PLX 87 IX 89 YYKG 94 KX 96 X 97 TKQLSNMIVX 107 SCX 110 CS

[0217] in,

[0218] X 3 Indicates D or W;

[0219] X 12 Indicates E or W;

[0220] X 40 Indicates E or Y;

[0221] X 50 Indicates P or S;

[0222] X 55 Indicates E or S;

[0223] X 66 Indicates K or H;

[0224] X 67 Indicates S or A;

[0225] X 68 Indicates H or R;

[0226] X 71 Indicates S or Q;

[0227] X 75 Indicates A, R or K;

[0228] X 81 Indicates S, E or K;

[0229] X 82 Indicates V or K;

[0230] X 84 Indicates A or E;

[0231] X 87 Indicates T or E;

[0232] X 89 Indicates K or T;

[0233] X 94 Indicates K, D, R or L;

[0234] X 96 Indicates L or K;

[0235] X 97 Indicates V or I;

[0236] X 107 Indicates Q, K or E;

[0237] X 110 Indicates T or R;

[0238] When X 66 Indicates H, X 67 Indicates A, X 68 When H is indicated, X 94 Not for R;

[0239] When X66 Indicates H, X 67 Indicates A, X 68 Indicates H, X 71 When X represents S, 94 When it is not N.

[0240] Preferably, in some examples, X 3 Indicates D.

[0241] Preferably, in some examples, X 12 Indicates E.

[0242] Preferably, in some examples, X 40 Indicates Y.

[0243] Preferably, in some examples, X 50 Indicates P.

[0244] Preferably, in some examples, X 55 Indicates E.

[0245] Preferably, in some examples, X 66 Indicates H.

[0246] Preferably, in some examples, X 67 Indicates A.

[0247] Preferably, in some examples, X 71 Indicates S.

[0248] Preferably, in some examples, X 75 Indicates K.

[0249] Preferably, in some examples, X 81 Indicates E.

[0250] Preferably, in some examples, X 82 Indicates K.

[0251] Preferably, in some examples, X 84 Indicates A.

[0252] Preferably, in some examples, X 87 Indicates T.

[0253] Preferably, in some examples, X 89 Indicates T.

[0254] Preferably, in some examples, X 94 Indicates K or R.

[0255] Preferably, in some examples, X 96 Indicates K.

[0256] Preferably, in some examples, X 97 Indicates V.

[0257] Preferably, in some examples, X 107 Indicates E.

[0258] Preferably, in some examples, X 110 Indicates R.

[0259] In some examples, the amino acid sequence of the recombinant polypeptide is shown in SEQ ID No. 19, SEQ ID No. 19:

[0260] GRKKRRQRRRPQGSGSALDTNYCFSSTETNCCVRPLYIDFEKDLGWKWIHEPKGYYANFCLGPCPPGWSLETQYSKVLALYHAX 68 NPSSSAKPCCVPEKLAPLTITYYKGX 94 KKVTKQLSNMIVESCRCS

[0261] in,

[0262] X 68 Indicates H or R;

[0263] X 94 Indicates K or R.

[0264] In some examples, the amino acid sequence of the recombinant polypeptide is any one of SEQ ID No. 20 to SEQ ID No. 30.

[0265] SEQ ID No.20:

[0266] GRKKRRQRRRPQGSGSALDTNYCFSSTETNCCVRPLYIDFEKDLGWKWIHEPKGYYAN FCLGPCPSGWSLRTQYSKVLALYKEHNPSSSSASPCCVPRVLAPLTIKYYKGKKLQTKQLSN MIVLSCTCS,

[0267] SEQ ID No.21:

[0268] GRKKRRQRRRPQGSGSALDTNYCFSSTETNCCVRPLYIDFEKDLGWKWIHEPKGYEAN FCLGPCPPGWSLETQYSKVLALYKSHNPQSSAAPCCVPKVLAPLEIKYYKGKKLVTKQLSN MIVQSCTCS,

[0269] SEQ ID No.22:

[0270] GRKKRRQRRRPQGSGSALDTNYCFSSTETNCCVRPLYIDFEKDLGWKWIHEPKGYYAN FCLGPCPPGWSLSTQYSKVLALYHSHNPSSSARPCCVPSVLEPLTITYYKGDKKITKQLSNMI VKSCTCS、

[0271] SEQ ID No.23:

[0272] GRKKRRQRRRPQGSGSALDTNYCFSSTETNCCVRPLYIDFEKDLGWKWIHEPKGYYAN FCLGPCPPGWSLRTQYSKVLALYHAHNPTSSARPCCVPRKLAPLTIEYYVGNKKVTKQLSN MIVQSCTCS、

[0273] SEQ ID No.24:

[0274] GRKKRRQRRRPQGSGSALDTNYCFSSTETNCCVRPLYIDFEKDLGWKWIHEPKGYYAN FCLGPCPPGWSLETQYSKVLALYHAHNPSSSAKPCCVPEKLAPLTITYYKGKKKVTKQLSN MIVESCRCS、

[0275] SEQ ID No.25:

[0276] GRKKRRQRRRPQGSGSALDTNYCFSSTETNCCVRPLYIDFEKDLGWKWIHEPKGYYAN FCLGPCPPGWSLRTQYSKVLALYKSHNPQSSAAPCCVPRSLSPLTITYYVGNTKITKQLSNMI VQACTCS、

[0277] SEQ ID No.26:

[0278] GRKKRRQRRRPQGSGSALWTNYCFSSTETNCCVRPLYIDFEKDLGWKWIHEPKGYYA NFCLGPCPPGWSLETQYSKVLALYHAHNPSSSAKPCCVPEKLAPLTITYYKGKKKVTKQLS NMIVESCRCS、

[0279] SEQ ID No.27:

[0280] GRKKRRQRRRPQGSGSALDTNYCFSSTWTNCCVRPLYIDFEKDLGWKWIHEPKGYYA NFCLGPCPPGWSLETQYSKVLALYHAHNPSSSAKPCCVPEKLAPLTITYYKGKKKVTKQLS NMIVESCRCS,

[0281] SEQ ID No.28:

[0282] GRKKRRQRRRPQGSGSALDTNYCFSSTETNCCVRPLYIDFEKDLGWKWIHEPKGYYAN FCLGPCPPGWSLETQYSKVLALYHARNPSSSAKPCCVPEKLAPLTITYYKGKKKVTKQLSN MIVESCRCS,

[0283] SEQ ID No.29:

[0284] GRKKRRQRRRPQGSGSALDTNYCFSSTETNCCVRPLYIDFEKDLGWKWIHEPKGYYAN FCLGPCPPGWSLETQYSKVLALYHARNPSSSAKPCCVPEKLAPLTITYYKGLKKVTKQLSN MIVESCRCS,

[0285] SEQ ID No.30:

[0286] GRKKRRQRRRPQGSGSALDTNYCFSSTETNCCVRPLYIDFEKDLGWKWIHEPKGYYAN FCLGPCPPGWSLETQYSKVLALYHARNPSSSAKPCCVPEKLAPLTITYYKGRKKVTKQLSN MIVESCRCS.

[0287] Further preferably, the amino acid sequence of the recombinant polypeptide is any one of SEQ ID No.24, SEQ ID No.28, and SEQ ID No.30.

[0288] In another aspect, the present invention provides a polynucleotide encoding the above polypeptide or recombinant polypeptide.

[0289] On the other hand, the present invention provides a composition comprising excipients and active ingredients, wherein the active ingredients include the above-mentioned polypeptide or a pharmaceutically acceptable salt thereof, or a recombinant polypeptide or a pharmaceutically acceptable salt thereof, or a polynucleotide; the excipients include at least one of a diluent, a filler, a binder, a wetting agent, an absorption enhancer, a surfactant, a lubricant and a stabilizer.

[0290] On the other hand, the present invention provides a use of the above-mentioned polypeptide or its pharmaceutically acceptable salt or recombinant polypeptide or its pharmaceutically acceptable salt or polynucleotide or pharmaceutical composition in the preparation of a drug for preventing and / or treating a disease associated with TGF-β.

[0291] In some embodiments, the use is to prevent hair loss and / or promote hair growth.

[0292] In some embodiments, the present invention provides a cosmetic composition for preventing hair loss or promoting hair growth, wherein the composition is selected from one or more of hair conditioners, hair essences, hair liquids, shampoos, hair conditioners, hair creams, hair waxes, hair aerosols, hair masks, hair nutrition packs, shampoo soaps, shampoo foams, hair oils, hair conditioners, hair dyes, hair curlers, and hair sprays.

[0293] The term "natural amino acid" refers to an amino acid that is genetically encoded by an organism itself. In certain preferred embodiments, the expression "natural amino acid" refers to the group consisting of alanine (A), arginine (R), aspartic acid (D), cysteine ​​(C), glutamine (Q), glutamic acid (E), histidine (H), isoleucine (I), glycine (G), aspartic acid (N), leucine (L), lysine (K), methionine (M), phenylalanine (F), co-peptide (P), serine (S), threonine (T), tryptophan (W), tyrosine (Y) and valine (V).

[0294] Site-Directed Mutagenesis is a molecular biology technique used to intentionally change specific amino acids in a protein sequence to study the effects of the amino acids on protein structure, function, or interaction with other molecules. Those skilled in the art know that the function of a small molecule polypeptide is closely related to its amino acid sequence, and changes in its sequence length, amino acid type, etc. may affect its function and effect. In this case, those skilled in the art cannot predict the effects of site-directed mutations on the function of a small molecule polypeptide sequence, nor can they predict the effects of site-directed mutations on the activity of a small molecule polypeptide sequence, such as CN106822865A and CN115385827A. Therefore, not all TGFβ polypeptide derivatives subjected to site-directed mutations can achieve the same effect as the polypeptide of the present application, that is, not all TGFβ polypeptide derivatives subjected to site-directed mutations can achieve the same effect as the polypeptide of the present application in preventing hair loss and / or promoting hair growth as effectively as the polypeptide of the present application.

[0295] The term "cell penetrating peptide (CPP)" is also known as "cell penetrating peptide", "protein translocation domain (PTD), "Trojan horse peptides" or "transduction peptide", etc., which refers to a peptide that can promote the transduction of various molecules (e.g., various macromolecules including proteins or nucleic acids; examples of the CPP include but are not limited to: (i) protein derived peptides: such as sequences derived from the gene controlling antennae (Antennapedia), such as pAntp (43-58); sequences derived from HIV-1, such as Tat derived peptides, such as amino acid residues 37-72, 37-60, 48-60 or 49-57 from TAT; (ii) model peptides: such as VT5; MAP; or arginine stretch, etc.; (iii) designed peptides: such as MPG; Transportan; Transportan 10; Pep-1; a peptide selected from KALA; or a peptide selected from Bulforin 2, and the like.

[0296] In addition, the CPP used in the conjugate of the present invention can also be selected from a polypeptide sequence having about 60, 70, 80, 90, 95, 99% or 100% sequence identity with any polypeptide sequence as described above, as long as the polypeptide sequence still retains its biological activity, i.e., promoting the cellular uptake of the isolated polypeptide of the present invention (or its variant) and / or promoting the isolated polypeptide of the present invention (or its variant) to pass through the blood-brain barrier.

[0297] Those skilled in the art can easily mutate the polypeptide of the present invention using known methods, such as site-directed mutagenesis or directed evolution. Those artificially modified sequences that have 90% or more identity with the polypeptide of the present invention have the same function as the polypeptide of the present invention, are derived from the nucleotide sequence of the present invention and are equivalent to the sequence of the present invention.

[0298] The term "alanine scanning mutagenesis" is a useful method for identifying residues or regions of a polypeptide that can be targeted for mutagenesis, as described in Cunningham and Wells (1989) Science, 244: 1081-1085. In this method, residues or target residue groups (e.g., charged residues such as arg, asp, his, lys, and glu) are identified and replaced with neutral or negatively charged amino acids (e.g., alanine or polyalanine) to determine whether the interaction of the polypeptide with its receptor is affected. Other substitutions can be introduced at amino acid positions that show functional sensitivity to the initial substitution. Alternatively or additionally, the crystal structure of the polypeptide-receptor complex can be used to identify contact points between the polypeptide and its receptor. Such contact residues and adjacent residues that are candidates for substitution can be targeted or eliminated. Variants can be screened to determine whether they have the desired properties. BRIEF DESCRIPTION OF THE DRAWINGS

[0299] Figure 1 This is a diagram showing the mass spectrometry detection result of the polypeptide shown in SEQ ID No.15;

[0300] Figure 2 This is a graph showing the cell proliferation promoting effect of the second generation human hair follicle dermal papilla cells (HFDPC);

[0301] Figure 3 This figure shows the cell proliferation promoting effect of the 4th passage human hair follicle dermal papilla cells (HFDPC). DETAILED DESCRIPTION

[0302] To make the purpose, technical scheme and advantages of the present invention clearer, the technical scheme of the present invention will be described in detail below. Obviously, the described embodiments are only a part of the embodiments of the present invention, rather than all the embodiments. Based on the embodiments in the present invention, all other embodiments obtained by ordinary technicians in the field without making creative work belong to the scope of protection of the present invention. In the following examples, unless otherwise specified, the raw materials used are commercially available.

[0303] Example 1 Synthesis of polypeptide

[0304] Peptide biological expression process

[0305] 1. Gene construction

[0306] The target polypeptide DNA sequence is designed as overlapping oligonucleotide primers and synthesized. The full-length target DNA sequence is obtained by PCR reaction, and the sequence is connected to the vector pET15b-sumo. The polypeptide sequence is located downstream of SUMO and fused with SUMO protein for expression. The connected vector is transferred into E. coli for culture, the plasmid is extracted and sequenced, and the sequencing results are analyzed to ensure consistency with the designed target sequence.

[0307] 2. Peptide Expression

[0308] The correctly constructed bacterial liquid was inoculated into SB medium, and 2 / 1000 ampicillin (mother solution concentration: 200 mg / mL) was added. After shaking culture at 37°C for 4 hours, the temperature was adjusted to 17°C and the shaking culture was continued for 1 hour. Then, IPTG was added to a final concentration of 1 mM, and shaking culture was carried out at 200 rpm overnight to induce expression.

[0309] 3. Peptide Purification

[0310] The overnight induced bacterial solution was poured into 288g urea and dissolved in a 60℃ water bath for 5min, then 6M NaOH was added for ultrasonic disruption (3S, 8S interval, 3 minutes, power 60%), and 17.7g 6M HCL was added for neutralization, and then incubated with 25mL nickel magnetic beads for 1 hour. After strong magnetic adsorption, the supernatant was removed and resuspended, washed 3 times and eluted with 500mM imidazole, and the eluent was added with SUMO protease and sheared at 4℃ overnight. After adding 8M guanidine hydrochloride to the shearing solution, the sample was loaded onto the equilibrated C18 reverse phase column (600mg), washed with 1% acetonitrile plus 1% acetic acid, and eluted with 50% acetonitrile, and the eluent was detected by SDS-PAGE electrophoresis.

[0311] 4. Peptide quality inspection

[0312] The molecular weight of the peptide was detected by mass spectrometry, and the purity of the peptide was detected by HPLC.

[0313] The polypeptides of the present invention can be synthesized by the above-mentioned biological expression method. The mass spectrometry results of the polypeptide shown in SEQ ID No. 15 are as follows Figure 1 shown.

[0314] Example 2 TGF-β cell level agonist activity detection

[0315] 1. Experimental materials:

[0316] name factory Part Number TGFβ1 GenScript Z03411 TGF-βReporterHEK-293cell Yoshiman Biology GM-C05346 384 White Transparent Cell Plate thermo 142762 One-lite Luciferase assay system Vazyme DD1203-04 Multifunctional cell imaging microplate reader Biotek Cytation5

[0317] 2. Experimental steps:

[0318] The day before the test, TGF-βReporter HEK293 cells were plated in a 384-well white transparent bottom cell plate at 6000 cells / well and 20μl / well, and incubated overnight for more than 16 hours. The next day, the cell plate that had been completely attached was removed, and the sample was diluted with Loading Buffer containing 0.1% BSA. The test sample, TGF-β1 protein and Buffer were added to the corresponding 384-well plate at 5uL / well (TGF-β1 protein as a positive control, 0.1% BSA Loading Buffer as a negative control), and placed in an incubator for 6 hours or overnight for about 20 hours. After incubation, the cell plate was removed for chemiluminescence detection on a multifunctional cell imaging microplate reader.

[0319] The agonist activity of the sample was calculated based on the agonist rate of TGF-β1 protein as 100%. The calculation formula is as follows:

[0320] Activation rate = 100% × (sample signal - buffer signal) / (TGF-β1 signal - buffer signal).

[0321] The concentration response curve of the polypeptide of the present invention on the activation of TGF-β receptor is drawn. The EC 50 The results are shown in Table 1. The polypeptides represented by SEQ ID No. 9, SEQ ID No. 13 and SEQ ID No. 15 have the strongest activation effect on TGF-β receptor.

[0322] Table 1 Response results of polypeptide concentration of the present invention

[0323]

[0324] Example 3 Human dermal papilla cell proliferation example

[0325] 1. Experimental materials and reagents:

[0326]

[0327]

[0328] name factory Part Number Cell Counting Kit-8 (CCK-8) GLPBIO GK10001 Transparent 96-well plate Guangzhou Jiete CAP011096

[0329] 2. Experimental Procedure

[0330] Human dermal papilla cells of the first four passages were selected and plated in a transparent 96-well plate coated with poly-L-lysine, with 10,000 cells / well, and placed at 37°C and 5% CO 2 In the incubator, after culturing for 24 hours, the original medium was discarded, and DMEM / F12 basic medium was added as a negative control, and the sample prepared in DMEM / F12 basic medium and TGF-β1 prepared in DMEM / F12 basic medium were added as a positive control. The volume of medium or drug in each well was 100 μL. After adding drugs, the cells were placed at 37°C and 5% CO 2 Incubate the cells in an incubator for 72 h. Add 10 μL of CCK8 reagent to each well and incubate at 37°C, 5% CO 2 Incubate in the incubator for 1 h and measure OD with an enzyme-labeled instrument 450 Absorbance value.

[0331] Proliferation rate calculation: Promotion rate (%) = (OD 450EXP -OD 450BLANK ) / (OD 450阴性对照 -OD 450BLANK )-1)×100%.

[0332] 3. Experimental results

[0333] The polypeptides of the present invention showed no toxicity or inhibitory effect on HFDPC cells. Figure 3 As shown, some of the calculation results are shown in Table 2. Compared with the blank group, the polypeptide of the present invention can significantly promote the proliferation of HFDPC cells; in the first 4 generations, the proliferation rate of each sample was relatively stable, and the proliferation rate at a higher concentration was 40%-70%, and the polypeptides shown in SEQ ID No.7, SEQ ID No.8 and SEQ ID No.11 had a relatively good concentration gradient effect.

[0334] Table 2 Calculation results of the proliferation rate of each sample

[0335]

[0336] The in vitro anti-hair loss experiment on mice of the present invention also confirms the effect of the polypeptide of the present invention in promoting hair growth and preventing or treating hair loss in vivo.

[0337] Finally, it should be noted that the above embodiments are only used to illustrate the technical solutions of the present invention, rather than to limit it. Although the present invention has been described in detail with reference to the aforementioned embodiments, those skilled in the art should understand that they can still modify the technical solutions described in the aforementioned embodiments, or make equivalent replacements for some of the technical features therein. However, these modifications or replacements do not deviate the essence of the corresponding technical solutions from the spirit and scope of the technical solutions of the embodiments of the present invention.

Claims

1. A polypeptide or a pharmaceutically acceptable salt thereof, characterized in that: The amino acid sequence of the polypeptide is shown in SEQ ID No. 1, SEQ ID No.1: ALX3TNYCFSSTX 12 TNCCVRPLYIDFEKDLGWKWIHEPKGYX 40 ANFCLGPCPX 50 GWSL X 55 TQYSKVLALYX 66 X 67 X 68 NPX 71 SSAX 75 PCCVPX 81 X 82 LX 84 PLX 87 IX 89 YYX 92 GX 94 X 95 X 96 X 97 TX 99 QLSNMIVX 107 X 108 CX 110 CS in, X3 represents D, W or Y; X 12 Indicates E or W; X 40 Indicates E or Y; X 50 Indicates P or S; X 55 Indicates R, E or S; X 66 Indicates K or H; X 67 It means E, S or A; X 68 Indicates H or R; X 71 Indicates S, Q or T; X 75 represents S, A, R or K; X 81 Indicates S, E, R or K; X 82 Indicates V, K, E or S; X 84 means A, E or S; X 87 Indicates T or E; X 89 represents K, T or E; X 92 Indicates K or V; X 94 Indicates K, D, N, R or L; X 95 Indicates K or T; X 96 Indicates L or K; X 97 Indicates Q, V or I; X 99 Indicates K or E; X 107 Indicates L, Q, K or E; X 108 Indicates S or A; X 110 Indicates T, R or V; When X 66 Indicates H, X 67 Indicates A, X 68 When H is indicated, X 94 Not for R; When X 66 Indicates H, X 67 Indicates A, X 68 Indicates H, X 71 When X represents S, 94 When it is not N.

2. The polypeptide or pharmaceutically acceptable salt thereof according to claim 1, characterized in that: The amino acid sequence of the polypeptide is shown in SEQ ID No. 2, SEQ ID No.2: ALX3TNYCFSSTX 12 TNCCVRPLYIDFEKDLGWKWIHEPKGYX 40 ANFCLGPCPX 50 GWSL X 55 TQYSKVLALYX 66 X 67 X 68 NPX 71 SSAX 75 PCCVPX 81 X 82 LX 84 PLX 87 IX 89 YYX 92 GX 94 X 95 X 96 X 97 TKQLSNMIVX 107 X 108 CX 110 CS in, X3 means D or W; X 12 Indicates E or W; X 40 Indicates E or Y; X 50 Indicates P or S; X 55 Indicates R, E or S; X 66 Indicates K or H; X 67 It means E, S or A; X 68 Indicates H or R; X 71 Indicates S, Q or T; X 75 represents S, A, R or K; X 81 Indicates S, E, R or K; X 82 Indicates V, K or S; X 84 means A, E or S; X 87 Indicates T or E; X 89 Indicates K, T or E; X 92 Indicates K or V; X 94 Indicates K, D, N, R or L; X 95 Indicates K or T; X 96 Indicates L or K; X 97 Indicates Q, V or I; X 107 Indicates L, Q, K or E; X 108 Indicates S or A; X 110 Indicates T, R; When X 66 Indicates H, X 67 Indicates A, X 68 When H is indicated, X 94 Not for R; When X 66 Indicates H, X 67 Indicates A, X 68 Indicates H, X 71 When X represents S, 94 When it is not N.

3. The polypeptide or pharmaceutically acceptable salt thereof according to claim 2, characterized in that: The amino acid sequence of the polypeptide is shown as SEQ ID No.3 or SEQ ID No.

4.

4. The polypeptide or pharmaceutically acceptable salt thereof according to any one of claims 1 to 3, characterized in that: The amino acid sequence of the polypeptide is any one of SEQ ID No.5 to SEQ ID No.

15.

5. A recombinant polypeptide or a pharmaceutically acceptable salt thereof, characterized in that: The recombinant polypeptide is obtained by coupling a cell penetrating peptide and the polypeptide according to claim 1 through a linker, wherein the coupling is covalently linked through an amide bond, and the linker comprises, for example, at least 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10 amino acid residues; the cell penetrating peptide is selected from any one or more of TAT peptide of HIV-1 virus reverse transcription activator protein, D-Tat, Antp peptide of Drosophila antennae homeotic protein, Pep-1 peptide, Pep-2 peptide, MPG peptide and RGD peptide.

6. The recombinant polypeptide or a pharmaceutically acceptable salt thereof according to claim 5, characterized in that: The amino acid sequence of the recombinant polypeptide is shown in SEQ ID No.16, SEQ ID No.16: SEQ ID No.16: GRKKRRQRRRPQGSGSALX3TNYCFSSTX 12 TNCCVRPLYIDFEKDLGWKWIHEPKGYX 40 ANFCLGPCPX 50 GWSLX 55 TQYSKVLALYX 66 X 67 X 68 NPX 71 SSAX 75 PCCVPX 81 X 82 LX 84 PLX 87 IX 89 YYX 92 GX 94 X 95 X 96 X 97 TX 99 QLSNMIVX 107 X 108 CX 110 CS in, X3 represents D, W or Y; X 12 Indicates E or W; X 40 Indicates E or Y; X 50 Indicates P or S; X 55 Indicates R, E or S; X 66 Indicates K or H; X 67 It means E, S or A; X 68 Indicates H or R; X 71 Indicates S, Q or T; X 75 represents S, A, R or K; X 81 Indicates S, E, R or K; X 82 Indicates V, K, E or S; X 84 means A, E or S; X 87 Indicates T or E; X 89 represents K, T or E; X 92 Indicates K or V; X 94 Indicates K, D, N, R or L; X 95 Indicates K or T; X 96 Indicates L or K; X 97 Indicates Q, V or I; X 99 Indicates K or E; X 107 Indicates L, Q, K or E; X 108 Indicates S or A; X 110 Indicates T, R or V; When X 66 Indicates H, X 67 Indicates A, X 68 When H is indicated, X 94 Not for R; When X 66 Indicates H, X 67 Indicates A, X 68 Indicates H, X 71 When X represents S, 94 When it is not N.

7. The recombinant polypeptide or a pharmaceutically acceptable salt thereof according to claim 6, characterized in that: The amino acid sequence of the recombinant polypeptide is shown as SEQ ID No.17, SEQ ID No.18 or SEQ ID No.

19.

8. The recombinant polypeptide or a pharmaceutically acceptable salt thereof according to any one of claims 5 to 7, characterized in that: The amino acid sequence of the recombinant polypeptide is any one of SEQ ID No.20 to SEQ ID No.

30.

9. A composition comprising an auxiliary material and an active ingredient, characterized in that: The active ingredient comprises the polypeptide or pharmaceutically acceptable salt thereof according to any one of claims 1 to 3, or the recombinant polypeptide or pharmaceutically acceptable salt thereof according to any one of claims 4 to 7, or the polynucleotide according to claim 8; the excipient comprises at least one of a diluent, a filler, a binder, a wetting agent, an absorption enhancer, a surfactant, a lubricant and a stabilizer.

10. Use of the polypeptide or pharmaceutically acceptable salt thereof according to any one of claims 1 to 4, the recombinant polypeptide or pharmaceutically acceptable salt thereof according to any one of claims 5 to 8, or the composition according to claim 9 for preventing and / or treating diseases associated with TGF-β, characterized in that: The application is prevention of hair loss and / or promotion of hair growth.

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