Polypeptide derivative, pharmaceutical composition containing polypeptide derivative, and use thereof and use method therefor

Abstract

A polypeptide derivative, which is characterized by comprising a polypeptide chain and a palmitic acid group modified on the polypeptide chain, and having a structure as represented by the following general formula I or general formula II: general formula I: C-X2, wherein in general formula I, X2 is absent, as well as any amino acid or any peptide segment, and C is modified with one palmitic acid group; and general formula II: X1-C-X3, wherein in general formula II, X1 and X3 are any amino acid or any peptide segment, and C is modified with two palmitic acid groups. The polypeptide derivative is capable of specifically binding to PIEZO2 and lowering a tactile threshold, thereby enhancing the effect of a PIEZO2-targeting agonist. A pharmaceutical composition containing the polypeptide derivative, when administered to mice, can effectively lower the mechanical pain threshold, thereby improving tactile sensitivity; and when administered to humans, the pharmaceutical composition can enhance the pleasure of human sexual activity.

Description

A polypeptide derivative, a pharmaceutical composition containing the polypeptide derivative, and its uses and methods of application. Technical Field

[0001] This invention relates to the technical field of peptides having an indeterminate number of amino acids, and more particularly to a polypeptide derivative that can act as an agonist of the tactile receptor Piezo2 and its applications. Background Technology

[0002] Touch plays an important role in daily life. The clinical manifestation of tactile hyposensitivity is a dulled sense of touch, making it difficult to avoid dangers that normal people can avoid, thus increasing vulnerability to injury. Current traditional treatments include behavioral and psychotherapy, with limited drug therapy options. Furthermore, tactile sensitivity is also related to sexual experience; lower sensitivity can negatively impact sexual harmony.

[0003] Regarding improving sexual experience, existing technology includes Chinese invention application CN201880098478.9 (publication number CN112839645A), entitled "Compositions and Methods for Treating Sexual Dysfunction and Enhancing Sexual Response and Pleasure," which discloses a drug that enhances human sexual response and sensitivity. Its active ingredients are cannabinoid derivatives and sexual response-enhancing components. Cannabinoid derivatives regulate dopamine secretion by activating the cannabinoid receptor CB1, thereby increasing sexual pleasure. While such drugs can enhance sexual pleasure in both men and women, cannabinoid derivatives are listed as Class I psychotropic drugs in my country, limiting their use and production processes, and posing a significant risk of addiction with long-term use. Therefore, the search for new drugs with novel mechanisms of action to enhance sexual pleasure is urgently needed.

[0004] Studies have shown that activating piezoelectric ion channel protein 2 (Piezo2) can enhance both the perception of mechanical force and sexual pleasure. Piezo2 is a mechanogated ion channel protein that can sense and respond to mechanical stimuli in cells, providing the molecular basis for the generation of tactile sensation. The Klaus corpuscle nerve endings in the sexual organs are enriched with mechanosensitive channels Piezo2, demonstrating that the presence of Piezo2 is essential for the sensitivity of the perineum caused by touch. Therefore, if Piezo2 can be activated with a certain chemical substance, the activator may be a potential drug for treating tactile hyposensitivity and enhancing sexual pleasure. However, no compound or pharmaceutical composition capable of activating Piezo2 and its application have yet been developed. Summary of the Invention

[0005] The first technical problem to be solved by the present invention is to provide a polypeptide derivative that can specifically bind to the Piezo2 channel, in light of the above-mentioned existing technology.

[0006] The second technical problem to be solved by the present invention is to provide a pharmaceutical composition containing the above-mentioned polypeptide derivative in view of the current state of the prior art.

[0007] The third technical problem to be solved by the present invention is to provide the use of the above-mentioned pharmaceutical composition in view of the current state of the prior art.

[0008] The fourth technical problem to be solved by the present invention is to provide a method for improving sexual pleasure by using the above-mentioned polypeptide derivatives or pharmaceutical compositions, in view of the current state of the prior art.

[0009] The fifth technical problem to be solved by the present invention is to provide a method for improving tactile sensitivity by applying the above-mentioned polypeptide derivatives or pharmaceutical compositions, in view of the current state of the prior art.

[0010] In a first aspect of the invention, a polypeptide is provided that specifically binds to the Piezo2 protein, the polypeptide being selected from the group consisting of:

[0011] (1) MDCGIMDACCESSDCLEICMECCGICFPS (SEQ ID NO: 8);

[0012] (2) CGIMDACCESSDCLEICMECCGICFPS (SEQ ID NO: 13);

[0013] (3)GIMDACCESSDCLEICMECCGICFPS (SEQ ID NO: 12);

[0014] (4) DACCESSDCLEICMECCGICFPS (SEQ ID NO: 11);

[0015] (5) CCESSDCLEICMECCGICFPS (SEQ ID NO: 10);

[0016] (6) CESADCLEICMECCGLCFSS (SEQ ID NO: 17);

[0017] (7) CESSDCLEICMECCGICFPS (SEQ ID NO: 9);

[0018] (8) SDCLEICMECCGICFPS (SEQ ID NO: 16);

[0019] (9) CLEICMECCGICFPS (SEQ ID NO: 5);

[0020] (10)LEICMECCGICFPS(SEQ ID NO:15);

[0021] (11)CLEICMECCGIC (SEQ ID NO:6);

[0022] (12) CMECCGICFPS (SEQ ID NO: 1);

[0023] (13) CLEICMECC (SEQ ID NO:7);

[0024] (14) CCGICFPS (SEQ ID NO:14);

[0025] (15)CMECCGIC(SEQ ID NO:2);

[0026] (16) CCGIC (SEQ ID NO:4);

[0027] (17)CMECC(SEQ ID NO:3).

[0028] In another preferred embodiment, the amino acids in the polypeptide are natural amino acids or non-natural amino acids.

[0029] In another preferred embodiment, the non-natural amino acids include: D-type amino acids, β-amino acids, and amino acids with side chain modifications.

[0030] In another preferred embodiment, the non-natural amino acids include: D-alanine (A), D-glutamic acid (E), D-aspartic acid (D), and D-serine (S).

[0031] In another preferred embodiment, a disulfide bond can be formed when two consecutive carbon atoms are present.

[0032] In a second aspect of the invention, a polypeptide derivative is provided, the polypeptide derivative comprising:

[0033] (a) the polypeptide described in the first aspect of the present invention; and

[0034] (b) A modifying group attached to an amino acid site selected from the group consisting of cysteine ​​(C), glutamic acid (E), serine (S), or a combination thereof.

[0035] In another preferred embodiment, the modifying group is attached to the cysteine ​​site of the polypeptide.

[0036] In another preferred embodiment, the modifying group is attached to one or more cysteine ​​sites of the polypeptide.

[0037] In another preferred embodiment, the polypeptide has one or more modifications at the same cysteine ​​site, preferably 1-5, more preferably 1-3.

[0038] In another preferred embodiment, the modification of the polypeptide by the modifying group gives the polypeptide the following function: improved tactile sensitivity.

[0039] In another preferred embodiment, the modifying group causes the polypeptide to have modifications selected from the group consisting of acylation, phosphorylation, glycosylation, cyclization, alkylation, or combinations thereof.

[0040] In another preferred embodiment, the polypeptide contains 0-5 types of modifications, more preferably 1-3, and more preferably 1-2.

[0041] In another preferred embodiment, the number of the modifying groups is 0-5, more preferably 1-3, and more preferably 1-2.

[0042] In another preferred embodiment, the modifying group used for acylation modification is selected from the group consisting of: myristoyl, palmitoyl, oleoyl, farnesyl, formyl, acetyl, propionyl, butyryl, malonyl, succinoyl, glutaryl, benzoyl, tetradecanoyl, scleroyl, biotinyl, fluorescent group, polyethylene glycol, or combinations thereof.

[0043] The modifying group is attached to the following positions of the polypeptide chain: N-terminus, C-terminus, side chain, or a combination thereof.

[0044] In another preferred embodiment, the side chain is a thiol side chain of cysteine.

[0045] In a third aspect of the present invention, a polypeptide derivative is provided, comprising a polypeptide chain and a modifying group modified on the polypeptide chain, having a structure shown in general formula I or general formula II below.

[0046] General Formula I: C-X2;

[0047] In general formula I, X2 can be none, any amino acid, or any peptide, and C can be modified with a modifying group.

[0048] General Formula II: X1-C-X3;

[0049] In general formula II, X1 and X3 are any amino acids or any peptide segments, and C is modified with m modifying groups.

[0050] Where C represents 1-5 amino acid sites in the peptide chain; m is an integer from 0 to 5.

[0051] In another preferred embodiment, X2 and X3 are polypeptides composed of 4-100 amino acid residues, more preferably 4-80 amino acid residues, more preferably 4-60 amino acid residues, and even more preferably 4-40 amino acid residues (e.g., polypeptides composed of 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, or 40 amino acid residues).

[0052] In another preferred embodiment, X1 is a polypeptide consisting of 1-50 amino acid residues, more preferably a polypeptide consisting of 1-30 amino acid residues, more preferably a polypeptide consisting of 1-20 amino acid residues, and even more preferably a polypeptide consisting of 1-10 amino acid residues (e.g., a polypeptide consisting of 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10 amino acid residues).

[0053] The modifying group is attached to the following positions of the polypeptide chain: N-terminus, C-terminus, side chain, or a combination thereof.

[0054] In another preferred embodiment, the side chain is a thiol side chain of cysteine.

[0055] In another preferred embodiment, the modifying group is selected from the group consisting of: myristoyl, palmitoyl, oleoyl, farnesyl, formyl, acetyl, propionyl, butyryl, malonyl, succinoyl, glutaryl, benzoyl, tetradecanoyl, scleroyl, biotinyl, fluorescent group, polyethylene glycol, or combinations thereof.

[0056] In another preferred embodiment, C is cysteine.

[0057] In another preferred embodiment, the polypeptide chain includes a polypeptide chain and modifying groups modified on the polypeptide chain, having the structure shown in general formula I or general formula II below;

[0058] General Formula I: C-X2;

[0059] In general formula I, X2 can be any amino acid or any peptide, and C is modified with a palmitic acid group.

[0060] General Formula II: X1-C-X3;

[0061] In formula II, X1 and X3 are any amino acids or peptides, and C is modified with two palmitic acid groups.

[0062] In another preferred embodiment, C is a cysteine ​​site.

[0063] In another preferred embodiment, the amino acid sequence of the polypeptide chain is selected from any one of SEQ ID NO:1-17, and polypeptides of their functional equivalent fragments, derivatives, and variants. In another preferred embodiment, the first cysteine ​​residue of the amino acid sequence of SEQ ID NO:1-6, SEQ ID NO:9, SEQ ID NO:10, and SEQ ID NO:13 is modified with a palmitic acid group;

[0064] The cysteine ​​residue at the third position of the amino acid sequences of SEQ ID NO:8 and SEQ ID NO:11 is modified with two palmitic acid groups.

[0065] The cysteine ​​residue at position 4 of the amino acid sequence of SEQ ID NO:1 is modified with two palmitic acid groups.

[0066] The cysteine ​​at position 6 of the amino acid sequence of SEQ ID NO:12 is modified with two palmitic acid groups.

[0067] In another preferred embodiment, the amino acid sequence of the polypeptide chain is selected from SEQ ID NO:1, and the first cysteine ​​residue of the polypeptide chain is modified with a palmitic acid group.

[0068] In another preferred embodiment, the polypeptide derivative (including the amino acid sequence and its modifying groups (Pal refers to palmitoyl group)) is selected from the group consisting of:

[0069] (M8): The following sequence and its modifications as shown in SEQ ID NO:8:

[0070] MDC(-2Pal)GIMDACCESSDCLEICMECCGICFPS;

[0071] (M13): The following sequence and its modifications as shown in SEQ ID NO:13:

[0072] Pal-CGIMDACCESSDCLEICMECCGICFPS;

[0073] (M20): The following sequence and its modifications as shown in SEQ ID NO:12:

[0074] GIMDACCESSDCLEICMEC(-Pal)CGICFPS;

[0075] (M12): The following sequence and its modifications as shown in SEQ ID NO:12:

[0076] GIMDAC(-2Pal)CESSDCLEICMECCGICFPS;

[0077] (M19): The following sequence and its modifications as shown in SEQ ID NO:11:

[0078] DACCESSDCLEICMEC(-Pal)CGICFPS;

[0079] (M11): The following sequence and its modifications as shown in SEQ ID NO:11:

[0080] DAC(-2Pal)CESSDCLEICMECCGICFPS;

[0081] (M10): The following sequence and its modifications as shown in SEQ ID NO:10:

[0082] Pal-CCESSDCLEICMECCGICFPS;

[0083] (M31): The following sequence and its modifications as shown in SEQ ID NO:17:

[0084] CESADCLEICMEC(-Pal)C(-Pal)GLC(-Pal)FSS;

[0085] (M32): The following sequence and its modifications as shown in SEQ ID NO:17:

[0086] CESADCLEIC(-Pal)MEC(-Pal)C(-Pal)GLC(-Pal)FSS;

[0087] (M33): The following sequence and its modifications as shown in SEQ ID NO:17:

[0088] CESADC(-Pal)LEIC(-Pal)MEC(-Pal)C(-Pal)GLC(-Pal)FSS;

[0089] (M9): The following sequence and its modifications as shown in SEQ ID NO:9:

[0090] Pal-CESSDCLEICMECCGICFPS;

[0091] (M18): The following sequence and its modifications as shown in SEQ ID NO:9:

[0092] CESSDCLEICMEC(-Pal)CGICFPS;

[0093] (M21): The following sequence and its modifications as shown in SEQ ID NO:9:

[0094] CESSDCLEICMECCGIC(-Pal)FPS;

[0095] (M22): The following sequence and its modifications as shown in SEQ ID NO:9:

[0096] CESSDCLEICMECC(-Pal)GICFPS;

[0097] (M23): The following sequence and its modifications as shown in SEQ ID NO:9:

[0098] CESSDCLEIC(-Pal)MECCGICFPS;

[0099] (M24): The following sequence and its modifications as shown in SEQ ID NO:9:

[0100] CESSDC(-Pal)LEICMECCGICFPS;

[0101] (M25): The following sequence and its modifications as shown in SEQ ID NO:9:

[0102] CESSDCLEICMECC(-Pal)GIC(-Pal)FPS;

[0103] (M26): The following sequence and its modifications as shown in SEQ ID NO:9:

[0104] CESSDCLEICMEC(-Pal)CGIC(-Pal)FPS;

[0105] (M27): The following sequence and its modifications as shown in SEQ ID NO:9:

[0106] CESSDCLEICMEC(-Pal)C(-Pal)GICFPS;

[0107] (M28): The following sequence and its modifications as shown in SEQ ID NO:9:

[0108] CESSDCLEICMEC(-Pal)C(-Pal)GIC(-Pal)FPS;

[0109] (M29): The following sequence and its modifications as shown in SEQ ID NO:9:

[0110] CESSDCLEIC(-Pal)MEC(-Pal)C(-Pal)GIC(-Pal)FPS;

[0111] (M30): The following sequence and its modifications as shown in SEQ ID NO:9:

[0112] CESSDC(-Pal)LEIC(-Pal)MEC(-Pal)C(-Pal)GIC(-Pal)FPS;

[0113] (M17): The following sequence and its modifications as shown in SEQ ID NO:16:

[0114] SDCLEICMEC(-Pal)CGICFPS;

[0115] (M4): The following sequence and its modifications as shown in SEQ ID NO:5:

[0116] Pal-CLEICMECCGICFPS;

[0117] (M16): The following sequence and its modifications as shown in SEQ ID NO:15:

[0118] LEICMEC(-Pal)CGICFPS;

[0119] (M5): The sequence shown in SEQ ID NO:6 and its modifications: Pal-CLEICMECCGIC; (M15): The sequence shown in SEQ ID NO:1 and its modifications:

[0120] CMEC(-Pal)CGICFPS;

[0121] (M7): The following sequence and its modifications as shown in SEQ ID NO:1:

[0122] CMEC(-2Pal)CGICFPS;

[0123] (M0): The following sequence and its modifications shown in SEQ ID NO:1: Pal-CMECCGICFPS;

[0124] (M6): The following sequence and its modifications shown in SEQ ID NO:7: Pal-CLEICMECC;

[0125] (M14): The following sequence and its modifications shown in SEQ ID NO:14: C(-Pal)CGICFPS;

[0126] (M1): The following sequence and its modifications shown in SEQ ID NO:2: Pal-CMECCGIC;

[0127] (M3): The following sequence and its modifications shown in SEQ ID NO:4: Pal-CCGIC;

[0128] (M2): The following sequence and its modifications shown in SEQ ID NO:3: Pal-CMECC.

[0129] This invention synthesizes corresponding polypeptide derivatives, denoted as M0 to M33 peptides, based on the amino acid sequences of polypeptides with SEQ ID NO: 1-17 and their corresponding modifying group sequences, collectively referred to as M peptides. The modifying group "(-Pal)" or "(-2Pal)" modifies the amino acid to the left of the group; the modifying group "Pal-" modifies the amino acid to the right of the group. "(-nPal)" or "(nPal-)" indicates that n palmitoyl (-Pal) groups are modified at a certain amino acid site. For example, "(-2Pal)" indicates that two palmitoyl groups are modified at a certain amino acid site.

[0130] In this invention, equivalent changes and modifications of polypeptide derivatives are all within the scope of protection. Specifically, this should also include polymers with structures shown in Formula I, such as dimers, trimers, etc. In this invention, polypeptide derivatives can also be modified or labeled by other common proteins, compounds, or polymers (e.g., phosphorylation, ubiquitination, PEG modification, etc.) to obtain fusion proteins or modified polypeptide derivatives.

[0131] In a fourth aspect of the invention, a probe is provided, which is obtained by fluorescent labeling a polypeptide derivative selected from the above.

[0132] In another preferred embodiment, the fluorescent labeling is performed by labeling with FITC dye.

[0133] In a fifth aspect of the invention, the use of the probe described in the fourth aspect of the invention is provided for: indicating the location of Piezo2 in a cell.

[0134] In another preferred aspect of the invention, the use of the probe described in the fourth aspect of the invention is provided for the preparation of a kit indicating the location of Piezo2 in cells.

[0135] In another preferred aspect of the invention, a kit is provided comprising the probe described in the fourth aspect of the invention.

[0136] In another preferred embodiment, in a sixth aspect of the invention, the use of the polypeptide derivative described in the second or third aspect of the invention is provided for the preparation of a medicament for treating sexual dysfunction with reduced tactile sensitivity.

[0137] In a seventh aspect of the invention, a pharmaceutical composition is provided comprising any polypeptide derivative or combination thereof described in the second or third aspect of the invention.

[0138] In another preferred embodiment, the pharmaceutical composition comprises any polypeptide derivative selected from the group consisting of, or a combination thereof:

[0139] (M8): The following sequence and its modifications as shown in SEQ ID NO:8:

[0140] MDC(-2Pal)GIMDACCESSDCLEICMECCGICFPS;

[0141] (M13): The following sequence and its modifications as shown in SEQ ID NO:13:

[0142] Pal-CGIMDACCESSDCLEICMECCGICFPS;

[0143] (M20): The following sequence and its modifications as shown in SEQ ID NO:12:

[0144] GIMDACCESSDCLEICMEC(-Pal)CGICFPS;

[0145] (M12): The following sequence and its modifications as shown in SEQ ID NO:12:

[0146] GIMDAC(-2Pal)CESSDCLEICMECCGICFPS;

[0147] (M19): The following sequence and its modifications as shown in SEQ ID NO:11:

[0148] DACCESSDCLEICMEC(-Pal)CGICFPS;

[0149] (M11): The following sequence and its modifications as shown in SEQ ID NO:11:

[0150] DAC(-2Pal)CESSDCLEICMECCGICFPS;

[0151] (M10): The following sequence and its modifications as shown in SEQ ID NO:10:

[0152] Pal-CCESSDCLEICMECCGICFPS;

[0153] (M31): The following sequence and its modifications as shown in SEQ ID NO:17:

[0154] CESADCLEICMEC(-Pal)C(-Pal)GLC(-Pal)FSS;

[0155] (M32): The following sequence and its modifications as shown in SEQ ID NO:17:

[0156] CESADCLEIC(-Pal)MEC(-Pal)C(-Pal)GLC(-Pal)FSS;

[0157] (M33): The following sequence and its modifications as shown in SEQ ID NO:17:

[0158] CESADC(-Pal)LEIC(-Pal)MEC(-Pal)C(-Pal)GLC(-Pal)FSS;

[0159] (M9): The following sequence and its modifications as shown in SEQ ID NO:9:

[0160] Pal-CESSDCLEICMECCGICFPS;

[0161] (M18): The following sequence and its modifications as shown in SEQ ID NO:9:

[0162] CESSDCLEICMEC(-Pal)CGICFPS;

[0163] (M21): The following sequence and its modifications as shown in SEQ ID NO:9:

[0164] CESSDCLEICMECCGIC(-Pal)FPS;

[0165] (M22): The following sequence and its modifications as shown in SEQ ID NO:9:

[0166] CESSDCLEICMECC(-Pal)GICFPS;

[0167] (M23): The following sequence and its modifications as shown in SEQ ID NO:9:

[0168] CESSDCLEIC(-Pal)MECCGICFPS;

[0169] (M24): The following sequence and its modifications as shown in SEQ ID NO:9:

[0170] CESSDC(-Pal)LEICMECCGICFPS;

[0171] (M25): The following sequence and its modifications as shown in SEQ ID NO:9:

[0172] CESSDCLEICMECC(-Pal)GIC(-Pal)FPS;

[0173] (M26): The following sequence and its modifications as shown in SEQ ID NO:9:

[0174] CESSDCLEICMEC(-Pal)CGIC(-Pal)FPS;

[0175] (M27): The following sequence and its modifications as shown in SEQ ID NO:9:

[0176] CESSDCLEICMEC(-Pal)C(-Pal)GICFPS;

[0177] (M28): The following sequence and its modifications as shown in SEQ ID NO:9:

[0178] CESSDCLEICMEC(-Pal)C(-Pal)GIC(-Pal)FPS;

[0179] (M29): The following sequence and its modifications as shown in SEQ ID NO:9:

[0180] CESSDCLEIC(-Pal)MEC(-Pal)C(-Pal)GIC(-Pal)FPS;

[0181] (M30): The following sequence and its modifications as shown in SEQ ID NO:9:

[0182] CESSDC(-Pal)LEIC(-Pal)MEC(-Pal)C(-Pal)GIC(-Pal)FPS;

[0183] (M17): The following sequence and its modifications as shown in SEQ ID NO:16:

[0184] SDCLEICMEC(-Pal)CGICFPS;

[0185] (M4): The following sequence and its modifications as shown in SEQ ID NO:5:

[0186] Pal-CLEICMECCGICFPS;

[0187] (M16): The following sequence and its modifications as shown in SEQ ID NO:15:

[0188] LEICMEC(-Pal)CGICFPS;

[0189] (M5): The sequence shown in SEQ ID NO:6 and its modifications: Pal-CLEICMECCGIC; (M15): The sequence shown in SEQ ID NO:1 and its modifications:

[0190] CMEC(-Pal)CGICFPS;

[0191] (M7): The following sequence and its modifications as shown in SEQ ID NO:1:

[0192] CMEC(-2Pal)CGICFPS;

[0193] (M0): The sequence shown in SEQ ID NO:1 and its modifications: Pal-CMECCGICFPS; (M6): The sequence shown in SEQ ID NO:7 and its modifications: Pal-CLEICMECC;

[0194] (M14): The following sequence and its modifications shown in SEQ ID NO:14: C(-Pal)CGICFPS;

[0195] (M1): The following sequence and its modifications shown in SEQ ID NO:2: Pal-CMECCGIC;

[0196] (M3): The following sequence and its modifications shown in SEQ ID NO:4: Pal-CCGIC;

[0197] (M2): The following sequence and its modifications shown in SEQ ID NO:3: Pal-CMECC.

[0198] In another preferred embodiment, the polypeptide derivative contains the amino acid sequence SEQ ID NO:1, and the first cysteine ​​residue of the polypeptide chain is modified with a palmitic acid group.

[0199] In another preferred embodiment, the dosage form of the pharmaceutical composition is a topical formulation.

[0200] In another preferred embodiment, the dosage form of the pharmaceutical composition is selected from the group consisting of liquid dosage forms, solid dosage forms, semi-solid dosage forms (e.g., gel dosage forms), or combinations thereof.

[0201] In another preferred embodiment, the dosage form of the pharmaceutical composition is selected from the group consisting of: ointments (e.g., creams or gels), injections, sprays, foams, suppositories, ointments, tablets, or combinations thereof.

[0202] In another preferred embodiment, the dosage form of the pharmaceutical composition is selected from the group consisting of: topical dosage forms, oral dosage forms, or combinations thereof.

[0203] In another preferred embodiment, the dosage form of the pharmaceutical composition is an injectable formulation.

[0204] In another preferred embodiment, the pharmaceutical composition is used for one or more uses selected from the group consisting of:

[0205] (i) Improve tactile sensitivity;

[0206] (ii) Enhance the pleasure of human sexual life;

[0207] (iii) Increase libido;

[0208] (iv) Increase vaginal secretions;

[0209] (v) Increase the frequency of orgasms;

[0210] (vi) Reduce insertion pain.

[0211] In an eighth aspect of the invention, use is provided for preparing a medicament, comprising the use of a polypeptide derivative described in the first aspect of the invention, a polypeptide derivative described in the second aspect of the invention, or a pharmaceutical composition described in the seventh aspect of the invention; said medicament is used for one or more uses selected from the group consisting of:

[0212] (i) Improve tactile sensitivity;

[0213] (ii) Enhance the pleasure of human sexual life;

[0214] (iii) Increase libido;

[0215] (iv) Increase vaginal secretions;

[0216] (v) Increase the frequency of orgasms;

[0217] (vi) Reduce insertion pain.

[0218] In a ninth aspect of the invention, a method for enhancing sexual pleasure is provided, the method comprising the steps of administering to a subject an effective amount of a polypeptide derivative of the second or third aspect of the invention, or a pharmaceutical composition of the seventh aspect of the invention.

[0219] In a tenth aspect of the invention, a method for improving tactile sensitivity is provided, the method comprising the steps of: administering to a test subject an effective amount of the polypeptide derivative of the second or third aspect of the invention, or the pharmaceutical composition of the seventh aspect of the invention.

[0220] In another preferred embodiment, the test subject is a human or a non-human mammal.

[0221] Compared with the prior art, the advantages of the present invention are as follows:

[0222] (1) The polypeptide derivatives in this invention can specifically bind to Piezo2 and reduce the tactile threshold, thereby enhancing the effect of Piezo2 target agonist.

[0223] (2) The polypeptide derivatives in this invention act on the Piezo2 channel, are non-addictive, and have high safety when used as drugs or as formulations.

[0224] (3) The probe in this invention can specifically bind to Piezo2 and can be used to indicate the location of Piezo2 in the cell;

[0225] (4) The pharmaceutical composition of the present invention uses the above-mentioned polypeptide derivatives. When applied to mice, it can effectively reduce the mechanical pain threshold and thereby improve tactile sensitivity; when applied to humans, it can improve the pleasure of human sexual life.

[0226] It should be understood that, within the scope of this invention, the above-described technical features of this invention and the technical features specifically described below (such as in the embodiments) can be combined with each other to form new or preferred technical solutions. Due to space limitations, they will not be described in detail here. Attached Figure Description

[0227] Figures 1a to 1d show the fluorescence colocalization of FITC-M peptide and Piezo2 fluorescent antibody in HeLa cells according to Example 2 of the present invention; wherein, Figure 1a is the red fluorescence labeling result of Piezo antibody; Figure 1b is the green fluorescence labeling result of FITC-M peptide; Figure 1c is the fluorescence signal overlap diagram of Figures 1a and 1b; and Figure 1d is a partial magnified view of Figure 1c.

[0228] Figures 2a and 2b show flow cytometry results of 293T cells and HeLa cells after treatment with peptide probes in Example 2 of the present invention; wherein, Figure 2a is a flow cytometry result of 293T cells after treatment with peptide probes; and Figure 2b is a flow cytometry result of HeLa cells after treatment with peptide probes.

[0229] Figure 3 shows a schematic diagram of the operation of the mechanical pain test of mouse foot in Example 4 of the present invention.

[0230] Figure 4 shows a bar chart of the 50% paw withdrawal response threshold of the experimental group and the control group in the mechanical pain test of mouse foot in Example 4 of the present invention; where Control is the control group and M-peptide is the experimental group; the relevant samples were subjected to a two-tailed t-test (n=8), p=0.017.

[0231] Figure 5 shows a bar chart of the 50% foot withdrawal response thresholds of peptides M0 to M13 in the experimental group of the mouse plantar mechanical pain test in Example 4 of the present invention.

[0232] Figure 6 shows the mouse paw withdrawal response ratio curve of Example 4 of the present invention.

[0233] Figures 7a and 7b show the experimental flowchart of the mouse mating behavior test in Example 4 of the present invention and the bar chart of the thrombus detection rate of the experimental group and the control group; wherein, Figure 7a is the experimental flowchart of the mouse mating behavior test; and Figure 7b is the bar chart of the thrombus detection rate of the experimental group and the control group in the mouse mating behavior test.

[0234] Figure 8 shows a bar chart illustrating the improvement in the sexual life of subjects by the Mo peptide and its pharmaceutical composition of Example 5 of the present invention.

[0235] Figure 9 shows the results of calcium ion hydrodynamic analysis.

[0236] Figure 10 shows the whole-cell patch-clamp recording data. Detailed Implementation

[0237] Through extensive and in-depth research and screening, the inventors unexpectedly discovered a class of polypeptide derivatives (M-peptides) that specifically bind to Piezo2. These derivatives significantly enhance tactile sensitivity and effectively improve sexual pleasure. In animal experiments, the inventors found that administration to mice increased mating rates and improved tactile sensitivity. In human experiments, the inventors found that application to the genital area enhanced sexual pleasure. Based on these findings, the present invention was completed.

[0238] The present invention will be further described in detail below with reference to the accompanying drawings and embodiments.

[0239] Existing drugs enhance sexual pleasure by activating the CB1 receptor and promoting dopamine secretion, but there is a risk of addiction with long-term use. This invention designs and develops peptide derivatives and pharmaceutical compositions that target and activate Piezo2, primarily to improve tactile sensitivity and enhance sexual pleasure.

[0240] In this invention, the inventors first designed a polypeptide derivative with the amino acid sequence shown in SEQ ID NO. 1-17 and modified with palmitic acid groups, based on the modification of one palmitic acid group with a cysteine ​​residue (hereinafter abbreviated as C) at the first position and two palmitic acid groups with cysteine ​​residues at the non-first and last positions. The inventors demonstrated through confocal microscopy and flow cytometry that the above-mentioned polypeptide derivative can specifically bind to Piezo2. Furthermore, in verifying the specific binding of the above-mentioned polypeptide derivative to Piezo2, the inventors also designed a probe capable of marking the location of Piezo2 in cells.

[0241] the term

[0242] To facilitate a clearer understanding of this disclosure, certain terms are first defined. As used herein, unless otherwise expressly specified herein, each of the following terms shall have the meaning given below. Other definitions are set forth throughout the application.

[0243] The term “about” can refer to a value or composition within an acceptable margin of error for a particular value or composition as determined by a person skilled in the art, depending in part on how the value or composition is measured or determined. For example, as used herein, the expression “about 100” includes all values ​​between 99 and 101.

[0244] As used herein, the terms “containing” or “including (comprise)” can be open-ended, semi-closed, or closed. In other words, the terms also include “consistently made of” or “composed of”.

[0245] As used herein, unless otherwise stated, any concentration range, percentage range, proportion range, or integer range shall be understood to include any integer value within the range and, where appropriate, its fractional value (e.g., one-tenth and one-hundredth of an integer).

[0246] As used herein, the term “and / or” refers to and covers any and all possible combinations of one or more of the related listed items.

[0247] As used in this article, "palmitoyl group," "palmitoyl group," "palmitoic acid group," and "palmitoyl group" have the same meaning and can be used interchangeably. The structural formula is: CH3(CH2). 14 C(O)- can be abbreviated as "-Pal" or "Pal-".

[0248] As used herein, "M" and "M peptide" have the same meaning, both referring to the polypeptide chain and modified polypeptide derivatives of the present invention. When a specific number is added after M (e.g., M1, M0, etc.), it refers to the corresponding polypeptide derivative containing a specific polypeptide chain and modified polypeptide derivatives.

[0249] Piezo2

[0250] Piezo2 is a mechanosensitive cation channel protein located on the cell membrane. Simply put, it is an extremely sophisticated "microscopic mechano-electrochemical converter" encoded by the cell itself. Its core function is to convert mechanical forces applied to the cell membrane (such as touch, tension, and pressure) into electrochemical signals, thereby allowing the organism to sense its own state and the external physical world.

[0251] The Piezo protein, shaped like a giant three-bladed propeller, is one of the largest known ion channels. This unique structure allows it to sense minute changes in cell membrane tension like a "nanospring." Unlike receptors that require chemical messenger (ligand) activation, Piezo2 is directly activated by physical forces, opening the channel and allowing calcium ions (Ca) to pass through. 2+ Sodium ions (Na) + The internal flow of these signals generates electrical signals (action potentials), which rapidly transmit information to the brain.

[0252] polypeptide derivatives

[0253] As used herein, the terms “peptide analogue,” “peptide derivative,” “derivative of the present invention,” and “M-peptide of the present invention” are used interchangeably and all refer to the peptide derivatives described in the second or third aspect of the present invention.

[0254] The present invention also provides a polypeptide derivative, which is described in the second or third aspect of the present invention.

[0255] Specifically, the present invention provides a polypeptide derivative, wherein the polypeptide derivative comprises:

[0256] (a) the polypeptide described in the first aspect of the present invention; and

[0257] (b) A modifying group attached to an amino acid site selected from the group consisting of cysteine ​​(C), glutamic acid (E), serine (S), or a combination thereof.

[0258] In another preferred embodiment, the modifying group is attached to the cysteine ​​site of the polypeptide.

[0259] In another preferred embodiment, the modifying group is attached to one or more cysteine ​​sites of the polypeptide.

[0260] In another preferred embodiment, the polypeptide has one or more modifications at the cysteine ​​sites, preferably 1-5, more preferably 1-3.

[0261] In another preferred embodiment, the modification of the polypeptide by the modifying group gives the polypeptide the following function: improved tactile sensitivity.

[0262] In another preferred embodiment, the modifying group causes the polypeptide to have modifications selected from the group consisting of acylation, phosphorylation, glycosylation, cyclization, alkylation, or combinations thereof.

[0263] In another preferred embodiment, the polypeptide contains 0-5 types of modifications, more preferably 1-3, and more preferably 1-2.

[0264] In another preferred embodiment, the number of the modifying groups is 0-5, more preferably 1-3, and more preferably 1-2.

[0265] In another preferred embodiment, the modifying group used for acylation modification is selected from the group consisting of: cardioyl, palmitoyl, scleroyl, oleoyl, farnesyl, acetyl, propionyl, butyryl, malonyl, succinyl, glutaryl, formyl, biotin, or combinations thereof.

[0266] In a preferred embodiment, the polypeptide derivative of the present invention comprises a polypeptide chain and a palmitic acid group modified on the polypeptide chain, having the structure shown in general formula I or general formula II below.

[0267] General Formula I: C-X2;

[0268] In general formula I, X2 can be any amino acid or any peptide, and C is modified with a palmitic acid group.

[0269] General Formula II: X1-C-X3;

[0270] In formula II, X1 and X3 are any amino acids or peptides, and C is modified with two palmitic acid groups.

[0271] In another preferred embodiment, the amino acid sequence of the polypeptide chain is selected from any one of SEQ ID No: 1-17;

[0272] In particular, the first cysteine ​​residue of the amino acid sequences SEQ ID NO:1-6, SEQ ID NO:9, SEQ ID NO:10, and SEQ ID NO:13 is modified with a palmitic acid group;

[0273] The cysteine ​​residue at the third position of the amino acid sequences of SEQ ID NO:8 and SEQ ID No:11 is modified with two palmitic acid groups;

[0274] The cysteine ​​residue at position 4 of the amino acid sequence of SEQ ID NO:1 is modified with two palmitic acid groups.

[0275] The cysteine ​​at position 6 of the amino acid sequence of SEQ ID NO:12 is modified with two palmitic acid groups.

[0276] The first cysteine ​​residue of the amino acid sequence of SEQ ID NO:1 is modified with a palmitic acid group.

[0277] In another preferred embodiment, the polypeptide derivative (including the amino acid sequence and its modifying groups (Pal refers to the palmitic acid group)) is selected from the group consisting of:

[0278] (M0): The following sequence and its modifications shown in SEQ ID NO:1: Pal-CMECCGICFPS;

[0279] (M1): The following sequence and its modifications shown in SEQ ID NO:2: Pal-CMECCGIC;

[0280] (M2): The following sequence and its modifications shown in SEQ ID NO:3: Pal-CMECC;

[0281] (M3): The following sequence and its modifications shown in SEQ ID NO:4: Pal-CCGIC;

[0282] (M4): The following sequence and its modifications as shown in SEQ ID NO:5:

[0283] Pal-CLEICMECCGICFPS;

[0284] (M5): The following sequence and its modifications shown in SEQ ID NO:6: Pal-CLEICMECCGIC;

[0285] (M6): The following sequence and its modifications shown in SEQ ID NO:7: Pal-CLEICMECC;

[0286] (M7): The following sequence and its modifications as shown in SEQ ID NO:1:

[0287] CMEC(-2Pal)CGICFPS;

[0288] (M8): The following sequence and its modifications as shown in SEQ ID NO:8:

[0289] MDC(-2Pal)GIMDACCESSDCLEICMECCGICFPS;

[0290] (M9): The following sequence and its modifications as shown in SEQ ID NO:9:

[0291] Pal-CESSDCLEICMECCGICFPS;

[0292] (M10): The following sequence and its modifications as shown in SEQ ID NO:10:

[0293] Pal-CCESSDCLEICMECCGICFPS;

[0294] (M11): The following sequence and its modifications as shown in SEQ ID NO:11:

[0295] DAC(-2Pal)CESSDCLEICMECCGICFPS;

[0296] (M12): The following sequence and its modifications as shown in SEQ ID NO:12:

[0297] GIMDAC(-2Pal)CESSDCLEICMECCGICFPS;

[0298] (M13): The following sequence and its modifications as shown in SEQ ID NO:13:

[0299] Pal-CGIMDACCESSDCLEICMECCGICFPS;

[0300] (M14): The following sequence and its modifications shown in SEQ ID NO:14: C(-Pal)CGICFPS;

[0301] (M15): The following sequence and its modifications as shown in SEQ ID NO:1:

[0302] CMEC(-Pal)CGICFPS;

[0303] (M16): The following sequence and its modifications as shown in SEQ ID NO:15:

[0304] LEICMEC(-Pal)CGICFPS;

[0305] (M17): The following sequence and its modifications as shown in SEQ ID NO:16:

[0306] SDCLEICMEC(-Pal)CGICFPS;

[0307] (M18): The following sequence and its modifications as shown in SEQ ID NO:9:

[0308] CESSDCLEICMEC(-Pal)CGICFPS;

[0309] (M19): The following sequence and its modifications as shown in SEQ ID NO:11:

[0310] DACCESSDCLEICMEC(-Pal)CGICFPS;

[0311] (M20): The following sequence and its modifications as shown in SEQ ID NO:12:

[0312] GIMDACCESSDCLEICMEC(-Pal)CGICFPS;

[0313] (M21): The following sequence and its modifications as shown in SEQ ID NO:9:

[0314] CESSDCLEICMECCGIC(-Pal)FPS;

[0315] (M22): The following sequence and its modifications as shown in SEQ ID NO:9:

[0316] CESSDCLEICMECC(-Pal)GICFPS;

[0317] (M23): The following sequence and its modifications as shown in SEQ ID NO:9:

[0318] CESSDCLEIC(-Pal)MECCGICFPS;

[0319] (M24): The following sequence and its modifications as shown in SEQ ID NO:9:

[0320] CESSDC(-Pal)LEICMECCGICFPS;

[0321] (M25): The following sequence and its modifications as shown in SEQ ID NO:9:

[0322] CESSDCLEICMECC(-Pal)GIC(-Pal)FPS;

[0323] (M26): The following sequence and its modifications as shown in SEQ ID NO:9:

[0324] CESSDCLEICMEC(-Pal)CGIC(-Pal)FPS;

[0325] (M27): The following sequence and its modifications as shown in SEQ ID NO:9:

[0326] CESSDCLEICMEC(-Pal)C(-Pal)GICFPS;

[0327] (M28): The following sequence and its modifications as shown in SEQ ID NO:9:

[0328] CESSDCLEICMEC(-Pal)C(-Pal)GIC(-Pal)FPS;

[0329] (M29): The following sequence and its modifications as shown in SEQ ID NO:9:

[0330] CESSDCLEIC(-Pal)MEC(-Pal)C(-Pal)GIC(-Pal)FPS;

[0331] (M30): The following sequence and its modifications as shown in SEQ ID NO:9:

[0332] CESSDC(-Pal)LEIC(-Pal)MEC(-Pal)C(-Pal)GIC(-Pal)FPS;

[0333] (M31): The following sequence and its modifications as shown in SEQ ID NO:17:

[0334] CESADCLEICMEC(-Pal)C(-Pal)GLC(-Pal)FSS;

[0335] (M32): The following sequence and its modifications as shown in SEQ ID NO:17:

[0336] CESADCLEIC(-Pal)MEC(-Pal)C(-Pal)GLC(-Pal)FSS;

[0337] (M33): The following sequence and its modifications as shown in SEQ ID NO:17:

[0338] CESADC(-Pal)LEIC(-Pal)MEC(-Pal)C(-Pal)GLC(-Pal)FSS.

[0339] In this invention, peptide derivatives were synthesized according to the amino acid sequences of SEQ ID NO:1-17 and their corresponding modifying groups, and were respectively denoted as M0 to M33 peptides, collectively referred to as M peptides.

[0340] In this invention, equivalent changes and modifications of polypeptide derivatives are all within the scope of protection. Specifically, this should also include polymers with structures shown in Formula I, such as dimers, trimers, etc. In this invention, polypeptide derivatives can also be modified or labeled by other common proteins, compounds, or polymers (e.g., phosphorylation, ubiquitination, PEG modification, etc.) to obtain fusion proteins or modified polypeptide derivatives.

[0341] More preferably, one or two amino acid residues on the polypeptide chain are replaced by one or more other amino acid residues. Even more preferably, one amino acid residue is replaced by another amino acid residue. Preferably, the replacement is homologous.

[0342] Homologous substitutions (the substitutions and replacements used in this article refer to the exchange between existing amino acid residues and optional residues) may occur, i.e., same-sex substitutions, such as basic substitution for basic, acidic substitution for acidic, polar substitution for polar, etc. Non-homologous substitutions may also occur, i.e., one residue is replaced by another, or optionally, non-natural amino acids, such as ornithine, diaminobutyric acid ornithine, ornithine-ornithine, pyridylalanine, thiophene-alanine, naphthylalanine, and phenylglycine, a more detailed list is provided below. More than one amino acid residue may be modified simultaneously. As used in this article, amino acids are classified according to the following categories: basic: H, K, R; acidic: D, E; nonpolar: A, F, G, I, L, M, P, V, W; polar: C, N, Q, S, T, Y.

[0343] Based on common knowledge in the art, those skilled in the art will know that in non-critical regions of a polypeptide, such as its ends, altering a few amino acid residues will not substantially change its biological activity; for example, appropriately substituting certain amino acids will not affect its activity. Therefore, those skilled in the art can implement such substitutions and ensure that the resulting molecule retains the desired biological activity.

[0344] Therefore, based on the teachings of this invention, it is obvious to those skilled in the art that mutants of the polypeptide chains of this invention can be obtained by mutating them while still possessing the desired function and activity. For example, it is known to those skilled in the art that adding or removing several amino acid residues, such as preferably 1-6, more preferably 1-3, and most preferably 1 amino acid residue, at either end of the polypeptide chain will not affect the function of the resulting mutant.

[0345] Therefore, this invention should include conserved mutants of the polypeptides of this invention. These conserved mutants can be generated by, for example, amino acid substitutions as shown in Table A below.

[0346] Table A

[0347] The polypeptide derivatives of the present invention can exist in the form of salts or esters, particularly pharmaceutically usable salts or esters.

[0348] Pharmaceutically usable salts of the polypeptide derivatives of the present invention include suitable acid addition salts or base salts thereof. They can form salts with strong inorganic acids, such as mineral acids (e.g., sulfuric acid, phosphoric acid, or hydrohalic acid); with strong organic carboxylic acids, such as unsubstituted or substituted (e.g., halogenated) alkylcarboxylic acids having 1 to 4 carbon atoms (e.g., acetic acid); with saturated or unsaturated dicarboxylic acids, such as oxalic acid, malonic acid, succinic acid, maleic acid, fumaric acid, phthalic acid, or terephthalic acid; with hydroxycarboxylic acids, such as ascorbic acid, glycolic acid, lactic acid, malic acid, tartaric acid, or citric acid; with amino acids, such as aspartic acid or glutamic acid; with benzoic acid; or with organic sulfonic acids, such as unsubstituted or substituted (e.g., halogenated) (C1-C4)-alkyl- or aryl-sulfonic acids (e.g., methanesulfonic acid or p-toluenesulfonic acid).

[0349] This invention also includes solvates of derivatives of this invention. The terms used in the claims include these forms. This invention also relates to various crystalline forms, polymorphs, and (anhydrous) aqueous forms of analogs of this invention. The pharmaceutical industry has established methods in which chemical compounds can be isolated in any of these forms by slightly modifying the purification and / or separation methods of the solvents used in the synthetic preparation of the compounds.

[0350] The present invention also includes derivatives of the invention in the form of prodrugs. Such prodrugs are typically derivatives of the invention, wherein one or more suitable groups are modified such that the modification is reversible upon administration to a human or mammalian subject. Although the reversal may also be achieved in vivo by administration of a second agent in conjunction with such a prodrug, such reversal is typically carried out by an enzyme naturally present in the subject's body. Examples of such modifications include esters (e.g., any of those described above), wherein the reversal can be carried out by an esterase. Other such systems are well known to those skilled in the art.

[0351] Pharmaceutical Composition

[0352] Based on the polypeptide derivative of the present invention, the present invention further provides a pharmaceutical composition that can improve tactile sensitivity or sexual pleasure, said pharmaceutical composition comprising a therapeutically effective amount of the polypeptide derivative of the present invention and a pharmaceutically acceptable carrier. Preferably, the pharmaceutical composition can also be used to: increase libido; increase vaginal secretions; increase the frequency of orgasms; and / or reduce penile pain.

[0353] As used herein, the term "effective dose" or "therapeutic effective dose" refers to a dose that is functional or active in humans and / or animals and is acceptable to humans and / or animals.

[0354] As used herein, the term "pharmaceuticalally acceptable" refers to a substance suitable for human and / or mammalian use without excessive adverse side effects (such as toxicity, irritation, and allergic reactions), i.e., a reasonable benefit / risk ratio. The term "pharmaceuticalally acceptable carrier" refers to a carrier used for the administration of therapeutic agents, including various excipients and diluents.

[0355] The preferred effective amount of the active ingredient (i.e., polypeptide derivative) described in this invention can be determined by those skilled in the art based on various factors (e.g., through clinical trials). These factors include, but are not limited to, the pharmacokinetic parameters of the active ingredient, such as bioavailability, metabolism, half-life, and route of administration.

[0356] The pharmaceutically acceptable carriers, effective amounts of active ingredients, and methods of administration described in this invention are well known to those skilled in the art.

[0357] The pharmaceutically acceptable carriers described in this invention include (but are not limited to): water, saline, liposomes, lipids, proteins, protein-antibody conjugates, peptides, cellulose, nanogels, or combinations thereof. The choice of carrier should be matched to the route of administration, as is well known to those skilled in the art.

[0358] The active ingredient of this invention can be administered or in combination with other drugs or preparations for the same purpose. In another preferred embodiment, the pharmaceutical composition further comprises one or more drugs or preparations that enhance sexual pleasure or lower the threshold of tactile sensation, or increase tactile sensitivity.

[0359] The pharmaceutical compositions of the present invention can be formulated into dosage forms such as injections, lyophilized formulations, nebulized inhalers, and topical preparations. The pharmaceutical compositions of the present invention can be delivered (administered) by any suitable method, including oral, parenteral, and topical methods. The pharmaceutical compositions of the present invention can also be administered by injection, i.e., intravenous, intramuscular, intradermal, subcutaneous, or intraperitoneal injection. Furthermore, the pharmaceutical compositions of the present invention can be administered transdermally. Transdermal administration via a local route can be formulated into medicated sticks, solutions, suspensions, emulsions, gels, creams, ointments, pastes, gels, paints, powders, and aerosols. In addition, the pharmaceutical compositions of the present invention can be actively administered to intradermal, subcutaneous, intramuscular, tissue, organ, and central nervous system sites via electrodes / electric fields / potential differences.

[0360] The pharmaceutical compositions of the present invention can be co-administered with another active agent. Co-administration includes administering the compound and active agent of the present invention within 0.5, 1, 2, 4, 6, 8, 10, 12, 16, 20, or 24 hours of each other. Co-administration also includes administering the compound and active agent of the present invention simultaneously, substantially simultaneously (e.g., within about 1, 5, 10, 15, 20, or 30 minutes of each other), or sequentially in any order. In some embodiments, co-administration can be accomplished by co-formulation, i.e., preparing a single pharmaceutical composition comprising both the active ingredient (peptide derivative) and the active agent of the present invention. In other embodiments, the active ingredient and active agent of the present invention can be formulated separately.

[0361] The present invention will be further illustrated below with reference to specific embodiments. It should be understood that these embodiments are for illustrative purposes only and are not intended to limit the scope of the invention. Experimental methods in the following embodiments, unless otherwise specified, are generally performed under conventional conditions, such as those described in Sambrook et al., Molecular Cloning: A Laboratory Manual (New York: Cold Spring Harbor Laboratory Press, 1989), or as recommended by the manufacturer. Unless otherwise stated, percentages and parts are weight percentages and parts by weight.

[0362] Example 1: Polypeptide Derivatives

[0363] The polypeptide derivative of this embodiment includes a polypeptide chain and a palmitic acid group modified on the polypeptide chain, and has the structure shown in general formula I or general formula II below;

[0364] General Formula I: C-X2,

[0365] In general formula I, X2 can be any amino acid or any peptide, and C is modified with a palmitic acid group.

[0366] General Formula II: X1-C-X3;

[0367] In formula II, X1 and X3 are any amino acids or peptides, and C is modified with two palmitic acid groups.

[0368] The amino acid sequence of the polypeptide chain is selected from any one of SEQ ID NO:1-13, wherein the first cysteine ​​residue of the amino acid sequence of SEQ ID NO:1-6, SEQ ID NO:9, SEQ ID NO:10, and SEQ ID NO:13 is modified with a palmitic acid group.

[0369] The cysteine ​​residue at the third position of the amino acid sequences of SEQ ID NO:8 and SEQ ID NO:11 is modified with two palmitic acid groups;

[0370] The fourth cysteine ​​residue in the amino acid sequence of SEQ ID NO:1 is modified with two palmitic acid groups.

[0371] The cysteine ​​at position 6 of the amino acid sequence of SEQ ID NO:12 is modified with two palmitic acid groups.

[0372] The amino acid sequences and their modifying groups of 34 polypeptide derivatives are shown below (Pal refers to the palmitic acid group):

[0373] SEQ ID NO:1(M0):Pal-CMECCGICFPS

[0374] SEQ ID NO:2(M1):Pal-CMECCGIC

[0375] SEQ ID NO:3(M2): Pal-CMECC

[0376] SEQ ID NO:4(M3): Pal-CCGIC

[0377] SEQ ID NO:5(M4):Pal-CLEICMECCGICFPS

[0378] SEQ ID NO:6(M5):Pal-CLEICMECCGIC

[0379] SEQ ID NO:7(M6):Pal-CLEICMECC

[0380] SEQ ID NO:1(M7):CMEC(-2Pal)CGICFPS

[0381] SEQ ID NO:8(M8):MDC(-2Pal)GIMDACCESSDCLEICMECCGICFPS

[0382] SEQ ID NO:9(M9):Pal-CESSDCLEICMECCGICFPS

[0383] SEQ ID NO:10(M10):Pal-CCESSDCLEICMECCGICFPS

[0384] SEQ ID NO:11(M11):DAC(-2Pal)CESSDCLEICMECCGICFPS

[0385] SEQ ID NO:12(M12):GIMDAC(-2Pal)CESSDCLEICMECCGICFPS

[0386] SEQ ID NO:13(M13):Pal-CGIMDACCESSDCLEICMECCGICFPS;

[0387] SEQ ID NO:14(M14):C(-Pal)CGICFPS;

[0388] SEQ ID NO:1(M15):CMEC(-Pal)CGICFPS;

[0389] SEQ ID NO:15(M16):LEICMEC(-Pal)CGICFPS;

[0390] SEQ ID NO:16(M17):SDCLEICMEC(-Pal)CGICFPS;

[0391] SEQ ID NO:9(M18):CESSDCLEICMEC(-Pal)CGICFPS;

[0392] SEQ ID NO:11(M19):DACCESSDCLEICMEC(-Pal)CGICFPS;

[0393] SEQ ID NO:12(M20):GIMDACCESSDCLEICMEC(-Pal)CGICFPS;

[0394] SEQ ID NO:9(M21):CESSDCLEICMECCGIC(-Pal)FPS;

[0395] SEQ ID NO:9(M22):CESSDCLEICMECC(-Pal)GICFPS;

[0396] SEQ ID NO:9(M23):CESSDCLEIC(-Pal)MECCGICFPS;

[0397] SEQ ID NO:9(M24):CESSDC(-Pal)LEICMECCGICFPS;

[0398] SEQ ID NO:9(M25):CESSDCLEICMECC(-Pal)GIC(-Pal)FPS;

[0399] SEQ ID NO:9(M26):CESSDCLEICMEC(-Pal)CGIC(-Pal)FPS;

[0400] SEQ ID NO:9(M27):CESSDCLEICMEC(-Pal)C(-Pal)GICFPS;

[0401] SEQ ID NO:9(M28):CESSDCLEICMEC(-Pal)C(-Pal)GIC(-Pal)FPS;

[0402] SEQ ID NO:9(M29): CESSDCLEIC(-Pal)MEC(-Pal)C(-Pal)GIC(-Pal)FPS;

[0403] SEQ ID NO:9(M30):

[0404] CESSDC(-Pal)LEIC(-Pal)MEC(-Pal)C(-Pal)GIC(-Pal)FPS;

[0405] SEQ ID NO:17(M31):CESADCLEICMEC(-Pal)C(-Pal)GLC(-Pal)FSS;

[0406] SEQ ID NO:17(M32):

[0407] CESADCLEIC(-Pal)MEC(-Pal)C(-Pal)GLC(-Pal)FSS;

[0408] SEQ ID NO:17(M33):

[0409] CESADC(-Pal)LEIC(-Pal)MEC(-Pal)C(-Pal)GLC(-Pal)FSS.

[0410] Specifically, the amino acid sequence of the polypeptide chain is preferably selected from SEQ ID NO:1, which has a palmitic acid group modified at the first cysteine ​​residue. The corresponding polypeptide derivatives are synthesized according to the amino acid sequences of SEQ ID NO:1-17 and their corresponding modified groups, and are denoted as peptides M0 to M33; collectively referred to as M peptides.

[0411] In this invention, equivalent changes and modifications of polypeptide derivatives are all within the scope of protection. Specifically, this should also include polymers with structures shown in Formula I, such as dimers, trimers, etc. In this invention, polypeptide derivatives can also be modified or labeled by other common proteins, compounds, or polymers (e.g., phosphorylation, ubiquitination, PEG modification, etc.) to obtain fusion proteins or modified polypeptide derivatives.

[0412] Example 2: Synthesis of peptide probes and experimental verification of their specific binding to Piezo2.

[0413] I. Synthesis of Peptide Probes

[0414] The polypeptide chain sequence of the polypeptide derivative in this embodiment can be selected from any one of SEQ ID NO:1-17. The polypeptide chain sequence of the polypeptide derivative selected in this embodiment is preferably SEQ ID NO:1. After synthesizing the M0 peptide modified with palmitic acid group according to the sequence, fluorescent labeling with fluorescein isothiocyanate (FITC) is used to obtain the polypeptide probe.

[0415] II. Confocal Experiment

[0416] To verify the specific binding of the synthesized peptide to Piezo2, HeLa cells highly expressing Piezo2 were used in this experiment. The cells were then conjugated with a 500 nM peptide probe and a fluorescently labeled Piezo2 antibody, respectively. The fluorescence colocalization of the peptide probe and the Piezo2 antibody in HeLa cells was observed using laser confocal microscopy. The experimental results are shown in Figures 1a to 1d. Red represents the fluorescence signal of the Piezo2 antibody, indicating the location of Piezo2 in the cell. Green represents the fluorescence signal of the peptide probe, indicating the location of Piezo2 in the cell. The figures show that the red fluorescence and green fluorescence can overlap well, with a colocalization coefficient M = 0.912 (M represents the Manders overlap coefficient, or MOC. When MOC is close to 1, it indicates that one signal almost completely overlaps with the other, meaning that the two signals are highly consistent in spatial location within the cell. When MOC is close to 0, it indicates that the two signals have almost no overlap, meaning that their positions in the cell structure are relatively independent). The peptide probe exhibits strong co-localization with Piezo2, and can be used to indicate the location of Piezo2 in cells, indicating that the peptide derivative of this embodiment can specifically bind to Piezo2.

[0417] III. Cell Flow Cytometry Experiment

[0418] To further verify that the polypeptide derivative synthesized in this embodiment can specifically bind to Piezo2, experiments were conducted using 293T cells that do not express Piezo2 and HeLa cells that highly express Piezo2. The two cell groups were treated with a 500 nM polypeptide probe to obtain experimental groups of 293T cells and HeLa cells. Within each cell population, control groups of 293T cells and HeLa cells were separated without probe treatment. The results were then analyzed by flow cytometry.

[0419] The results are shown in Figures 2a and 2b. Compared with the blank control group, 293T cells treated with FITC-M peptide did not show fluorescence shift, indicating that due to the lack of Piezo2 receptor, FITC-M peptide could not specifically bind to 293T cells. However, HeLa cells treated with FITC-M peptide showed obvious fluorescence shift, indicating that M peptide specifically binds to Piezo2.

[0420] Example 3 Pharmaceutical Composition

[0421] The pharmaceutical composition of this embodiment includes a polypeptide derivative M peptide or combination thereof, in which the amino acid sequence of the polypeptide chain selected from the polypeptide chain of Example 1 is selected from any of SEQ ID NO:1-17, and the amino acid sequence of the polypeptide chain in the polypeptide derivative is preferably SEQ ID NO:1, i.e., M0 peptide.

[0422] In this embodiment, the carrier or excipient of the pharmaceutical composition may be: sterile water, lauryl azone, solubilizer, propylene glycol, xanthan gum, hyaluronic acid, p-hydroxyacetophenone, 1,2-hexanediol, etc.

[0423] The specific preparation method of the drug composition is as follows: taking M0 peptide as an example, sterile water, lauryl azone, solubilizer, propylene glycol, xanthan gum, hyaluronic acid, p-hydroxyacetophenone, 1,2-hexanediol and other reagents are used as auxiliary reagents to prepare a gel formulation (the content of each excipient component varies from 0.5% to 90%), wherein the concentration of M0 peptide in the drug composition is 1-20 μM.

[0424] In addition, the dosage form of the drug composition can be an oral preparation, an injectable preparation or a topical preparation. In this embodiment, the dosage form is preferably a skin-applied preparation. The absorption principle of the drug composition in the skin-applied preparation is as follows: taking M0 peptide as an example, M0 peptide is absorbed into the body through the skin. Lauryl azone and propylene glycol in the formula can act as transdermal agents to help M0 molecules pass through the skin.

[0425] The dosage of the drug composition is designed as follows: 2 mL per person per use, in which the M peptide content is 10 μM. The actual dosage may be increased or decreased according to individual circumstances.

[0426] Example 4: Mouse Behavioral Testing

[0427] I. Mechanical Pain Test in Mice

[0428] The Von Frey (up-down) test (also known as the mouse plantar mechanical pain test) is used to measure the behavioral outcome of changes in the response of rodents to mechanical or thermal stimuli. The experimental procedure is as follows:

[0429] M0 to M33 peptides at 10 μM were formulated into topical formulations and applied to the soles of mice as the experimental group. Mice with a formulation without active peptides applied to their soles served as the control group. The other components and contents of the formulation without active peptides were the same as those in the experimental group.

[0430] Experimental mice (SPF mice, BALB / cJGpt) in both the experimental and control groups were gently placed in the test stand compartments and allowed to acclimatize for 1 hour. The experimental procedure is shown in Figure 3. Von Frey filaments were applied from the underside of the net to the sole of the mouse's hind paw, with a force starting at 0.6g. The test followed an up-down method to obtain maximum sensitivity until the filaments bent or the mouse responded.

[0431] A positive response is indicated by the animal readily withdrawing its paw upon application of a stimulus. Paw withdrawal responses in mice are observed; no response is recorded as a negative response "O", and a response (paw withdrawal or licking) is recorded as a positive response "X". If there is no response to the first filament stimulation, a filament with a slightly higher intensity is applied; if there is a response, a filament with a slightly lower intensity is applied, and so on, for a total of 5 measurements starting from the one with a response. This yields a sequence of "O" or "X" combinations. This sequence, along with the intensity (f) of the last filament, is input into a formula to calculate the mechanical pain threshold. The formula is: 50% paw withdrawal response threshold (g) = (10[Xf + kδ]) / 10000. Where Xf = log(f); δ is the average difference of each filament intensity after taking the logarithm, approximately 0.224 in this case; k is the value obtained by looking up the "X" and "O" sequences from a table. Data analysis uses the Up-Down Reader open-source program to effectively determine the 50% Von Frey threshold.

[0432] The experimental results are shown in Figures 4 and 5. The results indicate that the 14 polypeptide derivatives and their drugs from Example 1 can reduce the 50% withdrawal threshold (g) in mice to varying degrees, with M0 showing the most significant reduction. Therefore, the polypeptide derivatives synthesized in Example 1 (especially M0) can reduce the mechanical pain threshold, thereby improving tactile sensitivity. Thus, the polypeptide derivatives of Example 1 possess the function of a Piezo2 agonist.

[0433] II. Von Frey mouse paw withdrawal response ratio curve test

[0434] Experimental subjects: male or female SPF mice (BALB / cJGpt, 8-12 weeks old) from the same cohort;

[0435] Experimental reagents: Peptides M0 to M33 were prepared into topical formulations for skin application.

[0436] Experimental procedure: Mice in the experimental group were divided into 14 groups. Each group was treated with a skin-applied preparation of peptides M0 to M33 and applied to the soles of their feet. Mice in the control group were treated with a preparation without active peptides. Other components and contents were the same as those in the experimental group.

[0437] Before the experiment on the day of testing, mice were placed on a platform with an acrylic glass cover to acclimate for at least 30 minutes. Eight Von Frey microfilaments (0.008, 0.02, 0.04, 0.07, 0.16, 0.4, 0.6, 1.0, 1.4, 2 g) were applied, starting from the area with the lowest fiber strength (0.008 g) and progressing to the area with the highest fiber strength (2 g), for a total of three applications. Each experiment was repeated three times. Mice that retracted their paws in all applications were included in the statistical analysis. Sixteen mice were tested at each fiber strength level, and the percentage of mice retracting their paws was recorded. Each test was repeated at least three times within the same mouse cohort. A curve was plotted with the logarithm of fiber strength on the x-axis and the percentage of mice exhibiting paw-retraction in each group on the y-axis, resulting in Figure 6.

[0438] As shown in Figure 6, the experimental results indicate that there was a statistically significant difference in mouse datasets between the control group and the M0 peptide-treated group; there were 16 female mice. This suggests that M0 peptide can activate Piezo2 in mice, significantly increasing their sensitivity and the rate of foot withdrawal response.

[0439] III. Mouse Mating Behavior Test

[0440] Experimental environment: The test was conducted in a cage with a transparent plexiglass cover, which allowed for video recording and accurate behavioral scoring, and was carried out during a dark period;

[0441] Experimental subjects: asexual male and female SPF mice, BALB / cJGpt mice aged 8 to 16 weeks, with testosterone and follicle-stimulating hormone (FSH) levels in the mice within the same range as normal adult mice.

[0442] Experimental Procedure: On the first day, pretreatment was performed. Mice were grouped into cohorts of eight, with equal numbers of males and females. The experimental group received a topical formulation of peptides M0 to M33 applied to the gonads of both male and female mice. The control group received a formulation without active peptides. All other components and concentrations were the same as the experimental group. Before being placed in the same cages, male mice were housed separately and acclimatized to the laboratory for 1 hour. Female mice were housed in groups, and their estrus status was confirmed by vaginal irrigation and cytological examination before entering the test cages. After housing and confirmation, the mice were placed in the same cages, following the specific steps below:

[0443] The first step is to select one male mouse in estrus and one or two female mice between 4:30 and 5:00 a.m. on the second day and put them into the cage in the order of male first and female second.

[0444] The second step is overnight mating; mice usually mate at night, and a plug will form in the vagina after mating. This is the result of the coagulation gland secretion of the male mouse hardening when exposed to air.

[0445] The third step is to check the vaginal plugs between 8:30 and 9:00 AM on the second day after the couple is put together. Larger vaginal plugs tend to protrude outwards, while smaller ones are located inside the vagina or at the cervix, requiring the vaginal opening to be opened to see them. The vaginal plugs are used to determine whether mating was successful.

[0446] As shown in Figures 7a and 7b, the results indicate that the detection rate of thrombi in the experimental group was 75%, while that in the control group was 50%. The difference between the experimental group and the control group was significant, indicating that M-peptide and its drug composition can improve the mating rate of mice.

[0447] Example 5: Experiment on the improvement of human sexual life by M0 peptide preparation

[0448] Based on the skin-topical formulation containing M0 in Example 4, this example further develops a lubricant gel formulation by using the M0 peptide to prepare a gel solution for enhancing pleasure during human sexual activity.

[0449] In the initial phase, 22 physiologically healthy volunteers were recruited for the trial. The experimental dosage was 2 mL per person per use, containing 10 μM of M0 peptide. Five indicators were statistically analyzed using questionnaires: changes in sexual satisfaction, libido, sensitivity, sexual pleasure, and frequency of orgasm.

[0450] As shown in Figure 8, among the 22 samples collected, 63.3% of the subjects experienced an improvement in their sexual satisfaction, with significant differences in overall scores. This demonstrates that the M0 peptide and its drug from Example 1 have a significant effect on improving tactile sensitivity and sexual pleasure during sexual intercourse.

[0451] Example 6

[0452] To systematically verify the activation effect of the polypeptide derivative provided by this invention on the Piezo2 ion channel, this invention uses calcium ion hydrodynamic experiments for verification.

[0453] Human cervical cancer cells (HeLa) were used as a model in this experiment. HeLa cells in good growth condition were seeded in a specialized culture dish and incubated at 37°C in the dark for 45 minutes with a 5 μM concentration of the calcium ion-sensitive fluorescent dye Fluo-4 AM to allow the dye to fully enter the cells and hydrolyze into active Fluo-4.

[0454] After dye loading, cells were washed three times with preheated Hanks balanced salt solution to remove excess extracellular dye, and then incubated at 37°C in the dark for 15 minutes to ensure uniform intracellular dye distribution. Cells were then treated with the same concentrations of M0-M7 to prepare single-cell suspensions, which were then monitored in real-time using a flow cytometer at 37°C to capture the dynamic process of calcium ion influx.

[0455] The results are shown in the left figure of Figure 9. Among the several candidate peptide derivatives, only M0 can induce a rapid and significant calcium flow response. The fluorescence intensity increased sharply after drug administration, indicating that M0 can effectively increase the intracellular calcium ion concentration, suggesting that it has unique biological activity against a specific target.

[0456] To further investigate whether this calcium flow is specifically mediated by the Piezo2 channel, the inventors constructed a Piezo2 gene knockout (KO) HeLa cell line and set up wild-type (WT) cells as a control. Using the exact same Fluo-4 AM loading and flow cytometry assay, the calcium responses of the two cell types under M1 stimulation were compared.

[0457] As shown in the right panel of Figure 9, the peak calcium signal in Piezo2 KO cells was significantly reduced compared to WT cells, which strongly demonstrates that M0-induced calcium influx mainly depends on the function of Piezo2 channel protein.

[0458] Example 9

[0459] Based on the results of Example 8, after confirming that M0 can induce Piezo2-dependent calcium signaling, whole-cell patch-clamp recording was performed in order to directly record the activation of the channel at the electrophysiological level and further verify its specificity.

[0460] The experiment used three groups of cells: wild-type HeLa cells, wild-type cells treated with Yoda1 (a known Piezo1 agonist, used as an experimental control), and Piezo2 KO cells treated with M0.

[0461] The results are shown in Figure 10. The results indicate that in wild-type HeLa cells, M0 stimulation induces a large inward current similar to that induced by Yoda1 treatment; however, in Piezo2 KO cells, the peak current induced by M0 is significantly reduced. This result directly confirms functionally that M0 can specifically activate Piezo2 channels, generating typical inward cation currents.

[0462] Based on the experimental data from Examples 8 and 9, M0 can efficiently induce rapid and significant calcium ion flow in Piezo2 channel-dependent HeLa cells, demonstrating its unique biological activity. Furthermore, cell patch-clamp data confirm that M0 is an effective Piezo2 ion channel-specific chemoagonist, providing a powerful pharmacological tool for further in-depth research into the physiological and pathological functions of Piezo2 channels.

[0463] All documents mentioned in this invention are incorporated herein by reference as if each document were individually incorporated by reference. Furthermore, it should be understood that after reading the foregoing teachings of this invention, those skilled in the art can make various alterations or modifications to this invention, and these equivalent forms also fall within the scope defined by the appended claims.

Claims

1. A polypeptide derivative, characterized in that, It includes a polypeptide chain and modifying groups modified on the polypeptide chain, and has the structure shown in general formula I or general formula II below; General Formula I: C-X2; In general formula I, X2 can be none, any amino acid, or any peptide, and C can be modified with a modifying group. General Formula II: X1-C-X3; In general formula II, X1 and X3 are any amino acids or any peptide segments, and C is modified with m modifying groups. Where C represents 1-5 amino acid sites in the peptide chain; m is an integer from 0 to 5.

2. The polypeptide derivative according to claim 1, characterized in that, The amino acid sequence of the polypeptide chain is selected from any one of SEQ ID NO:1-17.

3. The polypeptide derivative according to claim 1, characterized in that, The modifying group is attached to the following positions of the polypeptide chain: N-terminus, C-terminus, side chain, or a combination thereof.

4. The polypeptide derivative according to claim 1, characterized in that, The modifying group is selected from the group consisting of: cardamomyl, palmitoyl, scleroyl, oleoyl, farnesyl, acetyl, propionyl, butyryl, malonyl, succinyl, glutaryl, formyl, biotin, or combinations thereof.

5. The polypeptide derivative according to claim 2, characterized in that, The polypeptide derivatives are selected from the following group: (M8): The following sequence and its modifications as shown in SEQ ID NO:8: MDC(-2Pal)GIMDACCESSDCLEICMECCGICFPS; (M13): The following sequence and its modifications as shown in SEQ ID NO:13: Pal-CGIMDACCESSDCLEICMECCGICFPS; (M20): The following sequence and its modifications as shown in SEQ ID NO:12: GIMDACCESSDCLEICMEC(-Pal)CGICFPS; (M12): The following sequence and its modifications as shown in SEQ ID NO:12: GIMDAC(-2Pal)CESSDCLEICMECCGICFPS; (M19): The following sequence and its modifications as shown in SEQ ID NO:11: DACCESSDCLEICMEC(-Pal)CGICFPS; (M11): The following sequence and its modifications as shown in SEQ ID NO:11: DAC(-2Pal)CESSDCLEICMECCGICFPS; (M10): The following sequence and its modifications as shown in SEQ ID NO:10: Pal-CCESSDCLEICMECCGICFPS; (M31): The following sequence and its modifications as shown in SEQ ID NO:17: CESADCLEICMEC(-Pal)C(-Pal)GLC(-Pal)FSS; (M32): The following sequence and its modifications as shown in SEQ ID NO:17: CESADCLEIC(-Pal)MEC(-Pal)C(-Pal)GLC(-Pal)FSS; (M33): The following sequence and its modifications as shown in SEQ ID NO:17: CESADC(-Pal)LEIC(-Pal)MEC(-Pal)C(-Pal)GLC(-Pal)FSS; (M9): The following sequence and its modifications as shown in SEQ ID NO:9: Pal-CESSDCLEICMECCGICFPS; (M18): The following sequence and its modifications as shown in SEQ ID NO:9: CESSDCLEICMEC(-Pal)CGICFPS; (M21): The following sequence and its modifications as shown in SEQ ID NO:9: CESSDCLEICMECCGIC(-Pal)FPS; (M22): The following sequence and its modifications as shown in SEQ ID NO:9: CESSDCLEICMECC(-Pal)GICFPS; (M23): The following sequence and its modifications as shown in SEQ ID NO:9: CESSDCLEIC(-Pal)MECCGICFPS; (M24): The following sequence and its modifications as shown in SEQ ID NO:9: CESSDC(-Pal)LEICMECCGICFPS; (M25): The following sequence and its modifications as shown in SEQ ID NO:9: CESSDCLEICMECC(-Pal)GIC(-Pal)FPS; (M26): The following sequence and its modifications as shown in SEQ ID NO:9: CESSDCLEICMEC(-Pal)CGIC(-Pal)FPS; (M27): The following sequence and its modifications as shown in SEQ ID NO:9: CESSDCLEICMEC(-Pal)C(-Pal)GICFPS; (M28): The following sequence and its modifications as shown in SEQ ID NO:9: CESSDCLEICMEC(-Pal)C(-Pal)GIC(-Pal)FPS; (M29): The following sequence and its modifications as shown in SEQ ID NO:9: CESSDCLEIC(-Pal)MEC(-Pal)C(-Pal)GIC(-Pal)FPS; (M30): The following sequence and its modifications as shown in SEQ ID NO:9: CESSDC(-Pal)LEIC(-Pal)MEC(-Pal)C(-Pal)GIC(-Pal)FPS; (M17): The following sequence and its modifications as shown in SEQ ID NO:16: SDCLEICMEC(-Pal)CGICFPS; (M4): The following sequence and its modifications as shown in SEQ ID NO:5: Pal-CLEICMECCGICFPS; (M16): The following sequence and its modifications as shown in SEQ ID NO:15: LEICMEC(-Pal)CGICFPS; (M5): The following sequence and its modifications shown in SEQ ID NO:6: Pal-CLEICMECCGIC; (M15): The following sequence and its modifications as shown in SEQ ID NO:1: CMEC(-Pal)CGICFPS; (M7): The following sequence and its modifications as shown in SEQ ID NO:1: CMEC(-2Pal)CGICFPS; (M0): The following sequence and its modifications shown in SEQ ID NO:1: Pal-CMECCGICFPS; (M6): The following sequence and its modifications shown in SEQ ID NO:7: Pal-CLEICMECC; (M14): The following sequence and its modifications shown in SEQ ID NO:14: C(-Pal)CGICFPS; (M1): The following sequence and its modifications shown in SEQ ID NO:2: Pal-CMECCGIC; (M3): The following sequence and its modifications shown in SEQ ID NO:4: Pal-CCGIC; (M2): The following sequence and its modifications shown in SEQ ID NO:3: Pal-CMECC; In this context, the palmitoyl group (-Pal) or (-2Pal) modification position is the amino acid site located to the left of the group; the palmitoyl group Pal- modification position is the amino acid site located to the right of the group.

6. A probe characterized by, It is obtained by fluorescent labeling of the polypeptide derivative as described in claim 1.

7. Use of the probe according to claim 6, characterized in that, Used to mark the location of Piezo2 in cells.

8. A pharmaceutical composition, characterized by, It includes any polypeptide derivative or combination thereof selected from those described in claim 1.

9. The pharmaceutical composition of claim 8, wherein, The dosage form of the pharmaceutical composition is a topical preparation.

10. Use of a polypeptide derivative according to claim 1 or a pharmaceutical composition according to claim 7, characterized in that, Used for the preparation of a medicine; wherein the medicine is used for one or more uses selected from the group consisting of: (i) Improve tactile sensitivity; (ii) Enhance the pleasure of human sexual life; (iii) Increase libido; (iv) Increase vaginal secretions; (v) Increase the frequency of orgasms; (vi) Reduce insertion pain.

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