Compounds as TGF-beta receptor agonist peptides and uses thereof

Abstract

The invention belongs to the field of polypeptide medicines, and particularly relates to a compound serving as a TGF-beta receptor agonist peptide and application of the compound. 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 in particular relates to a compound as a TGF-β receptor agonist peptide and its use. 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-β R) on the cell surface, which is divided into three subtypes: TGF-β RI (also known as ALK5), TGF-β RII, and TGF-β RIII. TGF-β forms a complex by binding to TGF-β RII with serine / threonine kinase activity on the cell membrane surface, and then forms a complex with TGF-β RI (also known as ALK5), thereby transmitting signals and activating corresponding signal 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-β RI (also known as ALK5) but retains the interaction with the receptors TGF-β RII and TGF-β RIII (see paragraphs [0015-0017] of the specification); existing TGF-β RI 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 thus are used for anti-tumor treatment.

[0006] Qixuan Wang's team at the University of California, Riverside, determined how TGF-β controls the cells in the hair follicle (including stem cells) to divide and form new cells, 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: One aspect of the present invention relates to a polypeptide, the amino acid sequence of which is shown in SEQ ID No. 1, SEQ ID No.1: ALX3TNYCFSSTETNCCVRPLYIDFEKDLGWKWIHEPKGYYANFCLGPCPPGWSLX 55 TQYSKVLALYHAHNPSSSAKPCCVPX 81 X 82 LAPLTIX 89 YYKG 94KX 96 VTX 99 QLSNMIVX 107 SCX 110 CS in, X3 represents D or Y; X 55 Indicates R or E; X 81 Indicates E or R; X 82 Indicates K or E; X 89 Indicates K or T; X 94 Indicates N or R; X 96 Indicates L or K; X 99 Indicates K or E; X 107 Indicates K or E; X 110 Indicates R or V.

[0008] In some examples, the amino acid sequence of the polypeptide is shown in SEQ ID No. 2, SEQ ID No. 3 or SEQ ID No. 4, SEQ ID No.2: ALDTNYCFSSTETNCCVRPLYIDFEKDLGWKWIHEPKGYYANFCLGPCPPGWSLRTQYSKVLALYHAHNPSSSAKPCCVPRELAPLTIKYYKGNKLVTEQLSNMIVKSCVCS, SEQ ID No.3: ALYTNYCFSSTETNCCVRPLYIDFEKDLGWKWIHEPKGYYANFCLGPCPPGWSLETQYSKVLALYHAHNPSSSAKPCCVPEKLAPLTITYYKGRKKVTKQLSNMIVESCRCS, SEQ ID No.4: ALDTNYCFSSTETNCCVRPLYIDFEKDLGWKWIHEPKGYYANFCLGPCPPGWSLETQYSKVLALYHAHNPSSSAKPCCVPEKLAPLTITYYKGRKKVTKQLSNMIVESCRCS.

[0009] 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 amino acids that play a key role in the activity of the polypeptide molecule.

[0010] 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.

[0011] Through site-directed mutagenesis technology, it was determined that E at position 25 of the polypeptide of the present invention is an amino acid essential for activity, further, "YIDFEKDLGW" at positions 20-30 is an amino acid essential for activity, further, "TNCCVRPLYIDFEKDLGWKWIHEPKGY" at positions 13-39 is an amino acid essential for activity, further, "TNYCFSSTETNCCVRPLYIDFEKDLGWKWIHEPKGYYANFCLGPCPPGWSL" at positions 3-54 and TQYSKVLALYHAHNPSSSAKPCCVP at positions 56-80 are amino acids essential for activity.

[0012] On the other hand, the present invention provides a recombinant polypeptide, wherein the recombinant polypeptide is obtained by coupling a cell penetrating peptide and a polypeptide shown in SEQ ID No. 1 via a linker, and the coupling is covalently linked via an amide bond.

[0013] In some examples, the linker comprises, for example, at least 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 amino acid residues.

[0014] 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.

[0015] Preferably, the linker is selected from GSGS.

[0016] 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.

[0017] In some examples, the cell penetrating peptide is selected from the group consisting of:

[0018] Preferably, in some examples, the cell penetrating peptide is selected from GRKKRRQRRRPQ.

[0019] 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.

[0020] 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.

[0021] 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.

[0022] In some examples, the cell penetrating peptide is connected to the N-terminus or C-terminus of the above polypeptide via a linker.

[0023] In some examples, the cell penetrating peptide is located at the N-terminus of the recombinant polypeptide.

[0024] In some examples, the cell penetrating peptide is located at the C-terminus of the recombinant polypeptide.

[0025] In some examples, the amino acid sequence of the recombinant polypeptide is shown in SEQ ID No. 5, SEQ ID No. 5: GRKKRRQRRRPQGSGSALX3TNYCFSSTETNCCVRPLYIDFEKDLGWKWIHEPKGYYANFCLGPCPPGWSLX 55 TQYSKVLALYHAHNPSSSAKPCCVPX 81 X 82 LAPLTIX 89 YYKG 94 KX 96 VTX 99 QLSNMIVX 107 SCX 110 CS in, X3 represents D or Y; X 55 Indicates R or E; X 81 Indicates E or R; X 82 Indicates K or E; X 89 Indicates K or T; X 94 Indicates N or R; X96 Indicates L or K; X 99 Indicates K or E; X 107 Indicates K or E; X 110 Indicates R or V.

[0026] In some examples, the amino acid sequence of the recombinant polypeptide is shown in SEQ ID No.6, SEQ ID No.7 or SEQ ID No.8, SEQ ID No.6: GRKKRRQRRRPQGSGSALDTNYCFSSTETNCCVRPLYIDFEKDLGWKWIHEPKGYYANFCLGPCPPGWSLRTQYSKVLALYHAHNPSSSAKPCCVPRELAPLTIKYYKGNKLVTEQLSNMIVKSCVCS, SEQ ID No.7: GRKKRRQRRRPQGSGSALYTNYCFSSTETNCCVRPLYIDFEKDLGWKWIHEPKGYYANFCLGPCPPGWSLETQYSKVLALYHAHNPSSSAKPCCVPEKLAPLTITYYKGRKKVTKQLSNMIVESCRCS, SEQ ID No.8: GRKKRRQRRRPQGSGSALDTNYCFSSTETNCCVRPLYIDFEKDLGWKWIHEPKGYYANFCLGPCPPGWSLETQYSKVLALYHAHNPSSSAKPCCVPEKLAPLTITYYKGRKKVTKQLSNMIVESCRCS.

[0027] On the other hand, the present invention provides a composition comprising excipients and an active ingredient, wherein the active ingredient comprises the above-mentioned polypeptide or recombinant polypeptide; the excipients comprise at least one of a diluent, a filler, a binder, a wetting agent, an absorption enhancer, a surfactant, a lubricant and a stabilizer.

[0028] In another aspect, the present invention provides a use of the above-mentioned polypeptide or recombinant polypeptide or composition in preventing and / or treating diseases associated with TGF-β.

[0029] In some embodiments, the use is to prevent hair loss and / or promote hair growth.

[0030] 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.

[0031] 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).

[0032] 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 the 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.

[0033] 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.

[0034] 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.

[0035] 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.

[0036] 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

[0037] Figure 1 This is a graph showing the HPLC test results of the polypeptide shown in SEQ ID No. 2; Figure 2 This is a diagram showing the mass spectrometry detection result of the polypeptide shown in SEQ ID No.2; Figure 3 This is a diagram showing the mass spectrometry detection result of the polypeptide shown in SEQ ID No.3; Figure 4 This is a graph showing the cell proliferation promoting effect of the second generation human hair follicle dermal papilla cells (HFDPC); Figure 5 This figure shows the cell proliferation promoting effect of the 4th passage human hair follicle dermal papilla cells (HFDPC). DETAILED DESCRIPTION

[0038] 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.

[0039] Peptide biological expression process

[0040] Example 1 Synthesis of polypeptide Peptide biological expression process 1. Gene construction 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.

[0041] 2. Peptide Expression The correctly constructed bacterial solution was inoculated into SB medium, and 2 / 1000 ampicillin (stock solution concentration: 200 mg / mL) was added. After shaking culture at 37°C for 4 hours, the temperature was adjusted to 17°C and shaking culture was continued for 1 hour. Then, IPTG was added to a final concentration of 1mM and shaking culture was carried out at 200 rpm overnight to induce expression.

[0042] 3. Peptide Purification The overnight induced bacterial solution was poured into 288g urea and dissolved in a 60℃ water bath for 5min, then ultrasonically disrupted with 6M NaOH (3S, 8S interval, 3min, 60% power), neutralized with 17.7g 6M HCL, 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 sheared with SUMO protease 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 and 1% acetic acid, and eluted with 50% acetonitrile. The eluent was detected by SDS-PAGE electrophoresis.

[0043] 4. Peptide quality inspection The molecular weight of the peptide was detected by mass spectrometry, and the purity of the peptide was detected by HPLC.

[0044] The polypeptides of the present invention can be synthesized by the above-mentioned biological expression method. The HPLC and mass spectrometry detection results of the polypeptides shown in SEQ ID No. 2 and SEQ ID No. 3 are as follows Figures 1 to 3 shown.

[0045] 1. Experimental materials:

[0046] 2. Experimental steps: The day before the test, TGF-β Reporter 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 completely adhered 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 incubation for 6 hours or overnight for about 20 hours. After incubation, the cell plate was removed and chemiluminescence detection was performed on a multifunctional cell imaging microplate reader. The test results calculated the agonist activity of the sample with the agonist rate of TGF-β1 protein as 100%, and the calculation formula is as follows: Excitation rate = 100% × (sample signal - buffer signal) / (TGF-β1 signal - buffer signal). 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. 2 and SEQ ID No. 3 have the strongest activation effect on TGF-β receptor.

[0047] Table 1 Response results of peptide concentration of the present invention

[0048] 1. Experimental materials and reagents:

[0049]

[0050] 2. Experimental Procedure The first four generations of human dermal papilla cells were selected and plated in a transparent 96-well plate coated with poly-lysine, 10,000 cells / well, and placed in a 37°C, 5% CO2 incubator. After culturing for 24 hours, the original medium was discarded, and DMEM / F12 basic medium was added as a negative control. The sample to be tested prepared in DMEM / F12 basic medium and TGF-β1 prepared in DMEM / F12 basic medium were added as positive controls. The volume of medium or drug in each well was 100 μL. After adding drugs, the cells were placed in a 37°C, 5% CO2 incubator for 72 hours. 10 μL CCK8 reagent was added to each well and incubated in a 37°C, 5% CO2 incubator for 1 hour, and the OD was detected by an enzyme reader. 450 Absorbance value.

[0051] Proliferation rate calculation: Promotion rate (%) = (OD 450 EXP -OD450 BLANK ) / (OD 450 阴性对照 -OD 450 BLANK )-1)×100%.

[0052] 3. Experimental results The polypeptides of the present invention showed no toxicity or inhibitory effect on HFDPC cells. Figure 4 , Figure 5 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 four generations, the proliferation rate of each sample was relatively stable, and the proliferation rate at higher concentrations was 40%-70%, and the polypeptide shown in SEQ ID No. 2 had a relatively good concentration gradient effect.

[0053] Table 2 Calculation results of proliferation rate of each sample

[0054] 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.

[0055] 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, characterized in that The amino acid sequence of the polypeptide is shown in SEQ ID No. 1, SEQ ID No.1: ALX3TNYCFSSTETNCCVRPLYIDFEKDLGWKWIHEPKGYYANFCLGPCPPGWSLX 55 TQYSKVLALYHAHNPSSSAKPCCVPX 81 X 82 LAPLTIX 89 YYKGX 94 KX 96 VTX 99 QLSNMIVX 107 SCX 110 CS in, X3 represents D or Y; X 55 Indicates R or E; X 81 Indicates E or R; X 82 Indicates K or E; X 89 Indicates K or T; X 94 Indicates N or R; X 96 Indicates L or K; X 99 Indicates K or E; X 107 Indicates K or E; X 110 Indicates R or V.

2. The polypeptide according to claim 1, characterized in that The amino acid sequence of the polypeptide is shown in SEQ ID No.2, SEQ ID No.3 or SEQ ID No.

4.

3. A recombinant polypeptide, 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.

4. The recombinant polypeptide according to claim 3, characterized in that The amino acid sequence of the recombinant polypeptide is shown in SEQ ID No. 5, SEQ ID No. 5: GRKKRRQRRRPQGSGSALX3TNYCFSSTETNCCVRPLYIDFEKDLGWKWIHEPKGYYANFCLGPCPPGWSLX55TQYSKVLALYHAHNPSSSAKPCCVPX 81 X 82 LAPLTIX 89 YYKGX 94 KX 96 VTX 99 QLSNMIVX 107 SCX 110 CS in, X3 represents D or Y; X 55 Indicates R or E; X 81 Indicates E or R; X 82 Indicates K or E; X 89 Indicates K or T; X 94 Indicates N or R; X 96 Indicates L or K; X 99 Indicates K or E; X 107 Indicates K or E; X 110 Indicates R or V.

5. The recombinant polypeptide according to claim 4, characterized in that The amino acid sequence of the recombinant polypeptide is shown as SEQ ID No.6, SEQ ID No.7 or SEQ ID No.

8.

6. A composition comprising an auxiliary material and an active ingredient, characterized in that: The active ingredient comprises the polypeptide according to claim 1 or 2 or the recombinant polypeptide according to any one of claims 3 to 5; 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.

7. Use of the polypeptide according to claim 1 or 2, the recombinant polypeptide according to any one of claims 3 to 5, or the composition according to claim 6 in 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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