Complex for improving drug stability and efficacy and use thereof

Abstract

The present invention provides a binary or ternary complex. The complex can improve the stability of a drug, and in particular, inhibit the transesterification reaction within drug molecules. The complex can also improve the efficacy formulation performance of the drug, improving the safety and effectiveness of the drug.

Description

A compound for improving drug stability and efficacy and its application

[0001] This application claims the benefit of priority of the following prior patent applications:

[0002] The applicant's prior application, patent application number 202410210343.4, filed with the State Intellectual Property Office of China on February 26, 2024, entitled "A composite for improving drug stability and efficacy and its application";

[0003] The entire contents of the above-mentioned prior patent applications are incorporated into this application by reference. Technical Field

[0004] The present invention belongs to the field of medicine, and in particular relates to a compound for improving drug stability and efficacy and application thereof. Background Art

[0005] Clascoterone (CLCT) is a new type of androgen receptor inhibitor. In August 2020, the U.S. FDA approved Cassiopea SpA's clacoterone cream (1% strength, trade name ) for the topical treatment of acne vulgaris in patients 12 years and older. It has also been disclosed that concentrations of 5.0% to 7.5% can be used to treat androgenetic alopecia (AGA), and 15% can be used to treat genital warts. However, this drug has poor stability and requires stringent storage conditions, requiring refrigeration. Its shelf life after opening is only two months, making it inconvenient for patients to use. Furthermore, the drug's high dosage concentration carries potential toxic side effects. Summary of the Invention

[0006] In order to improve the above technical problems, the present invention provides a complex and its application in improving drug stability and efficacy.

[0007] The present invention provides a complex consisting of [A] and [B] (a binary complex), or consisting of [A], [B], and [C] (a ternary complex);

[0008] wherein [A] is selected from long-chain fatty alcohols or polyols;

[0009] [B] and [C] are the same or different and are independently selected from terpene alcohols or organic acids.

[0010] According to an embodiment of the present invention, the long-chain fatty alcohols are C6-30 long-chain fatty alcohols, preferably C8-26 long-chain fatty alcohols, and more preferably C10-20 long-chain fatty alcohols.

[0011] According to an embodiment of the present invention, the long-chain fatty alcohols include long-chain saturated fatty alcohols and long-chain unsaturated fatty alcohols.

[0012] According to an embodiment of the present invention, the long-chain saturated fatty alcohol is a C6-30 long-chain saturated fat, preferably a C8-26 long-chain saturated fatty alcohol, and further preferably a C10-20 long-chain saturated fatty alcohol.

[0013] According to an embodiment of the present invention, the long-chain unsaturated fatty alcohol is a C6-30 long-chain unsaturated fatty alcohol, preferably a C8-26 long-chain unsaturated fatty alcohol, and further preferably a C10-20 long-chain unsaturated fatty alcohol.

[0014] According to an embodiment of the present invention, the long-chain fatty alcohols are selected from, for example, decyl alcohol, undecanol, lauryl alcohol, tridecanol, myristyl alcohol, pentadecanol, palmityl alcohol, heptadecanol, stearyl alcohol, oleyl alcohol, and linoleyl alcohol; preferably oleyl alcohol, linoleyl alcohol, and myristyl alcohol.

[0015] According to an embodiment of the present invention, the polyols are selected from polyols containing two or more hydroxyl groups, preferably from polyols containing two, three or four hydroxyl groups.

[0016] According to an embodiment of the present invention, the polyols are selected from C2-20 polyols, preferably C2-16 polyols, such as C2-10 polyols, C3-8 polyols, C4-6 polyols, and the like.

[0017] According to an embodiment of the present invention, the polyols are, for example, selected from glycerol, propylene glycol, 1,3-butanediol, and 1,2-pentanediol.

[0018] According to an embodiment of the present invention, terpene alcohols are terpene derivatives containing alcoholic hydroxyl groups, preferably C6-30 terpene derivatives containing alcoholic hydroxyl groups, and further preferably C10-20 terpene derivatives containing alcoholic hydroxyl groups, such as menthol, linalool, eucalyptol, santalol, nerol, geraniol, citronellol, terpineol, retinol, and ambroxol.

[0019] According to an embodiment of the present invention, the organic acid is selected from citric acid, citric acid, malic acid, lauric acid, geranic acid, and lactic acid.

[0020] According to an embodiment of the present invention, the molar ratio of [A] to [B] is 1:10-10:1, preferably 1:5-5:1, for example, 1:4.5, 1:4, 1:3.5, 1:3, 1:2.5, 1:2, 1:1.5, 1:1, 1.5:1, 2:1, 2.5:1, 3:1, 3.5:1, 4:1, 4.5:1.

[0021] According to an embodiment of the present invention, the molar ratio of [A], [B], and [C] is 1:10:10-10:1:1, preferably 1:5:5-5:1:1, for example 1:1:1.

[0022] According to an embodiment of the present invention, the molar ratio of [A] to [C] is 1:10-10:1, preferably 1:5-5:1, for example, 1:4.5, 1:4, 1:3.5, 1:3, 1:2.5, 1:2, 1:1.5, 1:1, 1.5:1, 2:1, 2.5:1, 3:1, 3.5:1, 4:1, 4.5:1.

[0023] According to an embodiment of the present invention, the molar ratio of [B] to [C] is 1:10-10:1, preferably 1:5-5:1, for example, 1:4.5, 1:4, 1:3.5, 1:3, 1:2.5, 1:2, 1:1.5, 1:1, 1.5:1, 2:1, 2.5:1, 3:1, 3.5:1, 4:1, 4.5:1.

[0024] According to an embodiment of the present invention, the complex is selected from any one of the following: a complex composed of oleyl alcohol and menthol, a complex composed of oleyl alcohol and citric acid, a complex composed of oleyl alcohol and lauric acid, a complex composed of oleyl alcohol and malic acid, a complex composed of oleyl alcohol and linalool, a complex composed of glycerol and malic acid, and a complex composed of myristyl alcohol, menthol, and linalool.

[0025] According to an embodiment of the present invention, the complex is selected from any one of the following: oleyl alcohol / menthol [OLO][MEO], oleyl alcohol / citric acid [OLO][CA], oleyl alcohol / lauric acid 2[OLO][LRA], oleyl alcohol / malic acid 4[OLO][MLA], oleyl alcohol / linalool [OLO][LNO], glycerol / malic acid 2[GLY]3[MLA], myristyl alcohol / menthol / linalool [MRO][MEO][LNO].

[0026] The present invention also provides a method for preparing the above-mentioned complex, which is as follows:

[0027] The composite is obtained by stirring [A] and [B] while keeping warm, or stirring [A], [B] and [C] while keeping warm.

[0028] According to an embodiment of the present invention, the preparation method of the composite is as follows:

[0029] [A] is stirred at a temperature maintained, and then [B] is added, or [B] and [C] are added, and the mixture is stirred at a temperature maintained, and cooled to obtain the composite.

[0030] According to an embodiment of the present invention, the temperature for heat preservation and stirring is 40°C-60°C, for example 50°C.

[0031] According to an embodiment of the present invention, the time of [A] heat preservation and stirring is 5 min-30 min, for example, 15 min.

[0032] According to an embodiment of the present invention, the time for continuing the heat preservation and stirring is 30 min-100 min, for example, 60 min.

[0033] According to an embodiment of the present invention, the cooling temperature is 0-40°C, such as room temperature.

[0034] The present invention provides application of the complex in improving drug stability.

[0035] According to an embodiment of the present invention, one end of the molecular structure of the drug is a hydroxyl group, and the other end is an alkyl ester group (eg, a C1-10 alkyl ester group, a C1-6 alkyl ester group, a C1-4 alkyl ester group, a C2 alkyl ester group).

[0036] According to an embodiment of the present invention, the hydroxyl group on the drug molecular structure can undergo an intramolecular transesterification reaction with an alkyl ester group.

[0037] According to an embodiment of the present invention, the chemical structure of the drug is shown in formula (I):

[0038] Wherein, M is selected from a drug core group;

[0039] L1 is selected from a bond, -C1-4 alkylene-carbonyl-, C1-4 alkylene-;

[0040] R1 is selected from C1-10 alkyl.

[0041] According to an embodiment of the present invention, L1 is selected from a bond, -C1-2 alkylene-carbonyl-, -C1-2 alkylene-; for example -CH2-CO- (carbonyl is connected to M).

[0042] According to an embodiment of the present invention, R1 is selected from C1-6 alkyl; preferably, R1 is selected from C1-4 alkyl; such as methyl, ethyl, propyl, butyl, etc.

[0043] According to an embodiment of the present invention, the chemical structure of the drug is shown in formula (II):

[0044] Wherein, M is selected from a drug core group.

[0045] According to an embodiment of the present invention, M is selected from steroidal drug nucleus groups.

[0046] According to an embodiment of the present invention, M is selected from the following groups:

[0047] Indicates the group attachment site;

[0048] R a 、R h 、R c 、R d 、R e The same or different, independently selected from H, C1-6 alkyl, halogen.

[0049] According to an embodiment of the present invention, R a 、R b The same or different groups are independently selected from C1-3 alkyl groups, such as methyl.

[0050] According to an embodiment of the present invention, R c 、R d 、R e are identical or different and are independently selected from H, halogen, for example H.

[0051] According to an embodiment of the present invention, the drug is a steroidal AR receptor antagonist, such as clacoterone.

[0052] According to an embodiment of the present invention, the complex improves drug stability by inhibiting the transesterification reaction within the drug molecule; preferably, the transesterification reaction is a propionate transesterification reaction.

[0053] The present invention also provides use of the complex in inhibiting transesterification reactions within drug molecules.

[0054] According to an embodiment of the present invention, the drug has the above definition.

[0055] The present invention also provides a use of the above-mentioned complex, wherein the complex is used to improve the efficacy of a drug.

[0056] According to an embodiment of the present invention, the drug is selected from steroid drugs, preferably steroid AR receptor antagonists; for example, clacoterone.

[0057] According to an embodiment of the present invention, the drug effect is selected from: antagonism of hormones (such as androgens, such as dihydrotestosterone DHT), oil-suppressing effect on cortical cells, anti-inflammatory effect, etc.

[0058] According to an embodiment of the present invention, the complex is used to enhance the antagonistic effect of clacoterone on androgens (such as dihydrotestosterone DHT).

[0059] According to an embodiment of the present invention, the complex is used to enhance the oil-suppressing effect of clacoterone on sebocytes (inhibiting the accumulation of lipids, such as triglycerides (TG)).

[0060] According to an embodiment of the present invention, the complex is used to enhance the anti-inflammatory effect of clacoterone.

[0061] The present invention also provides a composition or preparation, comprising the above-mentioned complex and a drug.

[0062] According to an embodiment of the present invention, the drug is selected from steroid drugs, preferably steroid AR receptor antagonists, such as clacoterone.

[0063] According to an embodiment of the present invention, in the composition or preparation, the mass ratio of the drug to the above-mentioned complex is 0.1%-20%:1%-30%, preferably 0.1%-10%:1%-20%, for example 1%:5%.

[0064] According to an embodiment of the present invention, the composition or preparation optionally further comprises a pharmaceutically acceptable excipient.

[0065] According to an embodiment of the present invention, the dosage form of the composition or preparation is selected from gels, ointments, creams, emulsions, foams, solutions, emulsions, suspensions, such as gels.

[0066] According to an embodiment of the present invention, the composition or formulation comprises the following components:

[0067] Clactophenone 0.1%-10% (e.g., 0.1%, 0.5%, 0.8%, 1%, 3%, 5%, 8%, 10%)

[0068] 1%-20% (e.g., 1%, 3%, 5%, 8%, 10%, 13%, 15%, 18%, 20%) of the above-mentioned complex.

[0069] According to an embodiment of the present invention, the composition or formulation comprises the following components:

[0070] The balance is water.

[0071] According to an embodiment of the present invention, the composition or preparation optionally further comprises a buffer solution; preferably, the antioxidant is selected from CA buffer solution.

[0072] The present invention also provides a method for preparing the above-mentioned composition or preparation, which is as follows:

[0073] The components are stirred and mixed to obtain the composition or preparation.

[0074] According to an embodiment of the present invention, the composition or preparation is prepared by: mixing propylene glycol and isopropyl alcohol, adding clacodone and the above-mentioned complex, and continuing to stir; adding water and HPC-H, and continuing to stir; and adjusting the pH to 4.2-4.9 to obtain the composition or preparation. According to an embodiment of the present invention, the HPC-H is sieved.

[0075] The present invention also provides use of the above composition or preparation in preparing a medicament for treating acne, alopecia (eg, androgenic alopecia), and genital warts. Beneficial effects

[0076] The present invention provides binary or ternary complexes that can improve drug stability, particularly inhibiting transesterification reactions within drug molecules. The complexes can also enhance drug efficacy and formulation performance, improving drug safety and effectiveness. BRIEF DESCRIPTION OF THE DRAWINGS

[0077] Figure 1 shows [OLO][MEO] 1 H NMR spectrum.

[0078] Figure 2 is the IR spectrum of [OLO][MEO].

[0079] FIG3 is a DSC graph of [OLO][MEO].

[0080] Figure 4 shows [OLO][CA] 1 H NMR spectrum.

[0081] Figure 5 shows 2[OLO][LRA] 1 H NMR spectrum.

[0082] Figure 6 shows the 4[OLO][MLA] 1 H NMR spectrum.

[0083] Figure 7 shows [OLO][LNO] 1 H NMR spectrum.

[0084] Figure 8 shows the relationship between 2[GLY]3[MLA] 1 H NMR spectrum.

[0085] Figure 9 shows [TEO][MEO][LNO] 1 H NMR spectrum.

[0086] FIG10 shows the enhancement of the AR receptor antagonistic effect of clacoterone by the complex [OLO][MEO], NS: p>0.05; *: p<0.05.

[0087] FIG11 is a picture of Nile Red staining of SZ95 sebocytes.

[0088] FIG12 shows the effect of the complex [OLO][MEO] on the anti-inflammatory and oil-suppressing effects of clacoterone.

[0089] FIG13 shows the effect of the complex [OLO][MEO] on the transdermal ability of clacoterone topical preparation.

[0090] FIG14 shows the effects of other compounds on the transdermal ability of clacoterone topical preparations. DETAILED DESCRIPTION

[0091] The technical solutions of the present invention will be described in further detail below with reference to specific embodiments. It should be understood that the following embodiments are merely illustrative and explanations of the present invention and should not be construed as limiting the scope of protection of the present invention. All technologies implemented based on the above content of the present invention are encompassed within the scope of protection that the present invention is intended to protect.

[0092] Unless otherwise specified, the raw materials and reagents used in the following examples are commercially available or can be prepared by known methods.

[0093] Example 1 Preparation of the composite

[0094] 1. Preparation of Oleyl Alcohol / Menthol [OLO] [MEO] Complex

[0095] prescription:

[0096] Process: Weigh oleyl alcohol in a round-bottom flask, keep warm at 50℃ and stir for 15 minutes; weigh menthol and add it to oleyl alcohol, continue to keep warm and stir for 60 minutes; cool to room temperature.

[0097] The hydrogen nuclear magnetic resonance (HNMR) spectrum of the oleyl menthol [OLO] [MEO] complex is shown in Figure 1 .

[0098] [OLO][MEO] H NMR spectrum multiplet data: 1 H NMR (400MHz, DMSO-d6) δ5.31 (m, 2H), 4.31 (m, 2H), 3.38 (m, 2H), 3.17 (tt, J=10.0, 4.8Hz, 1H), 2.21 (pd, J=7.0, 2.6Hz, 1H), 1 .98 (q, J=6.4Hz, 4H), 1.84 (m, 1H), 1.60 (dt, J=12.5, 3.0Hz, 1H), 1.52 (dq, J=12.4, 2.8Hz, 1H), 1.28 (m, 25H), 0.83 (m, 16H).

[0099] The infrared spectrum (IR) and differential scanning calorimetry (DSC) graphs are shown in Figures 2-3 respectively.

[0100] 2. Preparation of Oleyl Alcohol / Citric Acid [OLO][CA] Complex

[0101] prescription:

[0102] Process: Weigh oleyl alcohol in a round-bottom flask, keep warm at 50℃ and stir for 15 minutes; weigh citric acid and add it to oleyl alcohol, continue to keep warm and stir for 60 minutes; cool to room temperature.

[0103] The H NMR spectrum of the oleyl alcohol citrate [OLO][CA] complex is shown in Figure 4, and the multiplet data are as follows: 1 H NMR (400MHz, DMSO-d6) δ12.42 (s, 3H), 5.31 (t, J=5.0Hz, 2H), 5.19 (m, 1H), 3.35 (d, J=6.0Hz, 2H), 2.76 (dd, J=15 .1, 3.9Hz, 2H), 2.66 (m, 2H), 1.97 (q, J=6.0Hz, 4H), 1.39 (m, 2H), 1.26 (d, J=18.9Hz, 23H), 0.84 (d, J=6.3Hz, 3H).

[0104] 3. Preparation of Oleyl Alcohol / Lauryl 2[OLO][LRA] Complex

[0105] prescription:

[0106] Process: Weigh oleyl alcohol in a round-bottom flask, keep warm at 50℃ and stir for 15 minutes; weigh lauric acid and add it to oleyl alcohol, continue to keep warm and stir for 60 minutes; cool to room temperature.

[0107] The H NMR spectrum of the oleyl laurate 2[OLO][LRA] complex is shown in Figure 5, and the multiplet data are as follows: 1 H NMR (400MHz, DMSO-d6) δ11.97 (s, 1H), 5.30 (d, J=5.1Hz, 4H), 4.33 (d, J=5.2Hz, 2H), 3.36 (q, J=6.3, 5.5Hz, 4H), 2.17 (td, J =8.1, 7.6, 3.0Hz, 2H), 1.97 (q, J=6.5, 6.1Hz, 8H), 1.47 (m, 2H), 1.39 (t, J=6.6Hz, 4H), 1.23 (s, 60H), 0.84 (d, J=6.6Hz, 9H).

[0108] 4. Preparation of Oleyl Alcohol / Malic Acid 4[OLO][MLA] Complex

[0109] prescription:

[0110] Process: Weigh oleyl alcohol in a round-bottom flask, keep warm at 50℃ and stir for 15 minutes; weigh malic acid and add it to oleyl alcohol, continue to keep warm and stir for 60 minutes; cool to room temperature.

[0111] The H NMR spectrum of the oleylmalate 4[OLO][MLA] complex is shown in Figure 6, and the multiplet data are as follows: 1 H NMR (400MHz, DMSO-d6) δ12.42 (m, 2H), 5.56 (d, J = 66.8Hz, 1H), 5.29 (t, J = 5.2Hz, 8H), 4.33 (s, 5H), 3.35 (d, J = 6.3Hz, 8H), 2.60 (m, 1H), 2.47 (m, 1H), 1.95 (t, J=6.2Hz, 16H), 1.38 (t, J=6.7Hz, 8H), 1.25 (d, J=18.2Hz, 88H), 0.83 (d, J=6.7Hz, 12H).

[0112] 5. Preparation of Oleyl Alcohol / Linalool [OLO] [LNO] Complex

[0113] prescription:

[0114] Process: Weigh oleyl alcohol in a round-bottom flask, keep warm at 50℃ and stir for 15 minutes; weigh linalool and add it to oleyl alcohol, continue to keep warm and stir for 60 minutes; cool to room temperature to obtain the product.

[0115] The H NMR spectrum of the linalool [OLO] [LNO] complex is shown in Figure 7, and the multiplet data are as follows: 1 H NMR (400MHz, DMSO-d6) δ5.84 (dd, J=17.4, 11.0Hz, 1H), 5.31 (t, J=5.0Hz, 2H), 5. 10 (m, 2H), 4.93 (d, J = 10.7Hz, 1H), 4.44 (d, J = 2.8Hz, 1H), 4.33 (d, J = 5.2Hz, 1H), 3 .35(t, J=5.6Hz, 2H), 1.95(m, 6H), 1.62(s, 3H), 1.54(s, 3H), 1.38(dt, J=10.0, 5 .4Hz, 4H), 1.26 (d, J=18.0Hz, 22H), 1.13 (d, J=3.6Hz, 3H), 0.85 (t, J=6.0Hz, 3H).

[0116] 6. Preparation of Glycerol / Malic Acid 2[GLY]3[MLA] Complex

[0117] prescription:

[0118] Process: Weigh propylene glycol in a round-bottom flask, keep stirring at 50℃ for 15 minutes; weigh malic acid and add it to oleyl alcohol, continue to keep stirring for 60 minutes; cool to room temperature.

[0119] The H NMR spectrum of the glycerol malate 2[GLY]3[MLA] complex is shown in Figure 8, and the multiplet data are as follows: 1 H NMR (400MHz, DMSO-d6) δ12.51 (s, 6H), 5.74 (s, 2H), 4.38 (m, 4H), 4.06 (m, 2H), 3.45 (tdd, J=33 .3, 15.1, 5.7Hz, 8H), 2.81 (m, 3H), 2.71 (d, J=5.0Hz, 3H), 2.67 (d, J=5.2Hz, 3H), 2.54 (m, 3H).

[0120] 7. Preparation of Myristyl Alcohol / Menthol / Linalool [TEO][MEO][LNO] Complex

[0121] prescription:

[0122] Process: Weigh myristyl alcohol and menthol in a round-bottom flask, keep warm at 50°C and stir for 15 minutes; weigh linalool and add it to the flask, continue to keep warm and stir for 60 minutes; cool to room temperature to obtain the product.

[0123] The H NMR spectrum of the myristyl alcohol / menthol / linalool [TEO][MEO][LNO] complex is shown in Figure 9, and the multiplet data are as follows: 1 H NMR (400MHz, DMSO-d6) δ5.84 (ddt, J=16.1, 10.8, 2.7Hz, 1H), 5.10 (m, 2H), 4.93 (dd, J=10.5, 2.8Hz, 1H), 4.45 (t, J=2.7Hz, 1H), 4.33 (ddd, J=11.8, 6.1, 3.3Hz, 2H), 3.36 (t, J=5.8Hz, 2H), 3.14 (dq, J=1 0.4, 5.3Hz, 1H), 2.18 (q, J=7.1Hz, 1H), 1.92 (m, 2H), 1.82 (d, J=12.9Hz, 1H), 1.62 (d, J=3.9Hz, 4H), 1.53 (d, J=4.0Hz, 4H), 1.38 (m, 5H), 1.23 (m, 22H), 1.13 (t, J=2.6Hz, 3H), 0.85 (m, 12H), 0.73 (m, 4H).

[0124] Example 2 Effect of the Complex on the Stability of Clacotetone

[0125] Clascoterone (CLCT) is chemically unstable. During the formulation and storage process, the 17-propionate ester hydrolyzes to form 17α,21-dihydroxyprogesterone (impurity 1). This 17-propionate ester is then eliminated to form 21-hydroxypregna-4,16-diene-3,20-dione (impurity 2). The hydrolyzate then undergoes an esterification reaction with the 17-hydroxypropionate ester to form 17α-hydroxyprogesterone-21-propionate (impurity 3). Impurity 3 is the primary impurity produced during the degradation process, so the level of impurity 3 is used as an indicator of clacoterone's stability.

[0126] The structural formulas of clacotolone and impurity 3 are:

[0127] The effects of different complexes on the stability of clacodone were investigated: 20 mg of clacodone (CLCT) was weighed, 500 mg of propylene glycol was added to fully moisten the drug, and then 200 mg of different complexes were weighed and added. After vortex mixing, the mixture was placed under predetermined conditions (0 h, 25°C for 24 h, and 40°C for 24 h) to investigate changes in the content of impurity 3. The experimental results are shown in Table 1.

[0128] Table 1 Effect of the complex on the content of clacodone impurity 3

[0129] It can be seen from the test results in Table 1 that the addition of the complex can improve the stability of clacoterone and reduce the content of impurity 3 to varying degrees.

[0130] Example 3 Effect of Oleyl Alcohol / Menthol [OLO] [MEO] Complex on the Androgen Antagonism of Clactolide

[0131] AR receptors in hair follicle papilla cells, when affected by dihydrotestosterone (DHT), lead to cell shrinkage and a shortened growth phase, which is also the main pathogenesis of androgenic alopecia. Therefore, by culturing hair follicle papilla cells in vitro and detecting cell viability, it is possible to examine the strength of AR inhibitors in antagonizing the effects of androgens.

[0132] 1. Extraction and culture of hair follicle papilla cells (DPCs):

[0133] C57BL / 6 mice were killed by cervical dislocation, and after shaving excess dorsal hair, dorsal skin was removed. The fascia was carefully dissected using a scalpel and scissors, cut into long strips, and the dermis and subcutaneous fat were carefully dissected. The adipose tissue was digested overnight with 0.25% dispase, and residual hair shafts were carefully removed. The adipose tissue was further minced and digested with 0.2% collagenase for 4-6 hours, under microscopic observation. Digestion was terminated when the dermal sheath had become free cells and the dermal papilla had been freed from its root. The digestion solution was centrifuged and the supernatant discarded. The pellet was transferred to a dish containing DMEM and repeatedly washed. The dermal papilla and other cellular components were washed away, and the hair and fibrous tissue were discarded.

[0134] The washed solution was transferred to a low-speed centrifugation at 500 rpm for 3 minutes. Repeat this process 3-4 times, slowly aspirating and discarding the supernatant each time to remove free dermal sheath cells. The bottom suspension contains dermal papilla cells. After further centrifugation to remove the supernatant, add DMEM medium supplemented with 10% fetal bovine serum and culture in a cell culture incubator. Change the medium every 2 days. When the cell density reaches 90%, passage or freeze the cells. Use the third generation cells for subsequent experiments.

[0135] 2. Cell Culture and Experimental Grouping

[0136] Take the culture medium of dermal papilla cells in the logarithmic growth phase and set the grouping as follows:

[0137] Control: negative control, routine culture without addition;

[0138] A: positive control, 2 μM DHT was added;

[0139] B: 2 μM DHT and 1 μM clacodone were added;

[0140] C: 2 μM DHT and 2 μM clacodone were added;

[0141] D: 2 μM DHT, 1 μM clacodone, and 2 μM [OLO][MEO] were added;

[0142] 3. Detection of DPCs cell viability by CCK-8 assay

[0143] The density of DPCs was adjusted, and 100 μL (about 2000 cells) of cell suspension was inoculated into a 96-well plate per well. A blank well without cells was also set up. Cells were cultured for 48 h according to the experimental grouping conditions. After the cell culture was completed, 10 μL of CCK-8 solution was added to each well and incubated for another 2 h. The absorbance at 450 nm was measured using a microplate reader.

[0144] Cell viability (%) = (OD sample -OD blank ) / (OD control-OD blank )*100%.

[0145] 4. Test results

[0146] The results are shown in Figure 10. The viability of DPCs in group A decreased significantly (cell viability was 65.39%), indicating that dihydrotestosterone (DHT) at this concentration significantly inhibited the proliferation of dermal papilla cells; the cell viability of DPCs in groups B and C improved (cell viability was 81.56% and 94.11%, respectively), and the cell viability of group C was well maintained, indicating that clacoterone can exert an antagonistic effect on androgens, and the 2μM concentration is more effective than the 1μM concentration; the viability of DPCs in group D was higher (cell viability was 91.99%), which was better than that of group B (p<0.05) and equivalent to that of group C (p>0.05), indicating that the addition of [OLO][MEO] significantly enhanced the efficacy of clacoterone, allowing it to exert the effect of a 2μM concentration at a concentration of 1μM.

[0147] Example 4 Effect of Oleyl Alcohol / Menthol [OLO] [MEO] Complex on the Anti-inflammatory and Oil-Suppressing Effects of Clactoterone

[0148] 1. Cell Culture and Experimental Grouping

[0149] SZ95 sebaceous cells were selected for the study and cultured in DMEM medium containing 10% fetal bovine serum, 100 mg / mL streptomycin, and 100 U / mL penicillin, and the cells were maintained at 37°C and 5% CO2.

[0150] Sebocytes were inoculated into 6-well plates. The experimental groups were as follows, with 6 replicate wells in each group.

[0151] Control: negative control group, with normal saline;

[0152] A(T+NS): positive control group, containing 100 nM linoleic acid + 100 mM testosterone; plus normal saline;

[0153] B (T+CLCT): T1 group, containing 100 nM linoleic acid + 100 mM testosterone; plus 1 mM clacodone;

[0154] C (T+CLCT+complex): T2 group, containing linoleic acid 100nM + testosterone 100mM; plus clacoterone 1mM and [OLO][MEO] 2mM.

[0155] 2. Nile Red staining observation

[0156] Three replicate wells were obtained, the culture supernatant was discarded, and Nile Red (5 μg / mL) was added for staining. After staining, trypsin was added for 2 minutes to terminate the trypsin reaction and the culture medium was renewed to obtain a cell suspension. The results were observed using an inverted fluorescence microscope. The experimental results are shown in Figure 11.

[0157] As shown in Figure 11, under the action of linoleic acid and testosterone, the lipid accumulation of SZ95 epithelial cells increased significantly; clacoterone can antagonize the effect of testosterone and inhibit lipid accumulation; after adding [OLO][MEO], the effect of clacoterone was further enhanced, and the lipid accumulation level of sebaceous cells was close to that of the blank control group.

[0158] 3. Determination of inflammatory factor IL-6

[0159] After 24 h of culture, the cell culture medium was aspirated and centrifuged at 12000 RPM at 4°C for 5 min. The supernatant was used for subsequent IL-6 detection. The IL-6 ELISA kit (Elabscience) was returned to temperature. 100 μL of standard working solution or sample was added to the corresponding plate wells and incubated at 37°C for 90 min. After discarding the liquid in the plate, 100 μL of biotinylated antibody working solution was immediately added and incubated at 37°C for 60 min. The liquid in the plate was discarded and the plate was washed three times. 100 μL of HRP enzyme conjugate working solution was added to each well and incubated at 37°C for 30 min. The liquid in the plate was discarded and the plate was washed five times. 90 μL of substrate solution was added to each well and incubated at 37°C for about 15 min. 50 μL of stop solution was added to each well. The reading was immediately carried out at a wavelength of 450 nm and the data was processed.

[0160] 4. Determination of triglyceride TG content

[0161] Sebocytes were collected and added to the extract in the triglyceride (TG) fluorescence test kit. The cells were disrupted by ultrasonication for 10 minutes using a cell ultrasonic disruptor. The cells were centrifuged at 12,000 rpm for 10 minutes at 4°C, and the supernatant was collected. The supernatant was divided into two parts, one for determining the total amount of protein in the sample and the other for determining the TG content.

[0162] First, use the BCA protein concentration assay kit (Elabscience) to determine the total amount of protein in the sample for subsequent calculation of TG content. The specific steps are as follows: prepare BCA working solution according to the instructions; prepare standard solutions with concentrations of 0, 0.1, 0.2, 0.3, 0.4, 0.6, 0.8, and 1.0 mg / mL; set up standard wells and sample wells on the well plate, with n = 3 replicates for each sample; add 20 μL of standard solution or sample solution to the well; add 200 μL of BCA working solution to each well in the air and shake to mix for 20 seconds; cover the well with a film and react at 37°C for 30 minutes; measure the OD value at 562 nm using a microplate reader. The protein content was calculated using the following formula:

[0163] Total protein concentration (mg / mL) = (ΔA 562 -b)÷a×f;

[0164] ΔA 562 : Sample OD value - blank OD value (OD value when the standard concentration is 0);

[0165] a: slope of the standard curve;

[0166] b: intercept of standard curve;

[0167] f: dilution factor.

[0168] The triglyceride (TG) fluorescence assay kit (Elabscience) was then used to determine and calculate the TG content in the sample. The specific steps were as follows: Before testing, the reagents in the kit were equilibrated to room temperature, and the working solution was prepared according to the instructions. Standard solutions were prepared at concentrations of 0, 6, 12, 15, 18, 21, 24, and 30 μmol / mL. Standard and sample wells were set on the plate, with n = 3 replicates for each sample. 20 μL of the standard solution or sample solution was added to each well. 200 μL of the colorimetric working solution was added to each well and mixed thoroughly. The samples were incubated at 37°C for 5 minutes, and the fluorescence measurement was performed using the microplate reader with the parameters Ex / Em = 535 nm / 587 nm. The triglyceride content was calculated using the following formula:

[0169] Triglyceride content (μmol / gprot) = (ΔF-b) ÷ a ÷ C pr ×f;

[0170] ΔF: sample fluorescence value - blank fluorescence value (fluorescence value when the standard concentration is 0);

[0171] a: slope of the standard curve;

[0172] b: intercept of standard curve;

[0173] C pr : total protein concentration, gprot / L;

[0174] f: dilution factor.

[0175] The IL-6 and TG levels of the Control group were used as the benchmark, and the IL-6 and TG levels of each group were compared with those of the Control group to obtain the relative concentrations of each group. The test results are shown in Figure 12:

[0176] Comparison of group A and control group showed that under the stimulation of linoleic acid and testosterone, lipids (triglycerides TG) in SZ95 sebocytes accumulated in large quantities and produced more inflammatory factors; comparison of group A and group B showed that clacoterone had a significant efficacy, antagonizing the effects of testosterone, and the lipid accumulation in sebaceous cells and the expression of inflammatory factors were significantly inhibited; comparison of group B and group C showed that the additional addition of [OLO][MEO] could significantly enhance the oil-suppressing effect of clacoterone on sebaceous cells (p<0.05).

[0177] Example 5 Effect of Oleyl Alcohol / Menthol [OLO] [MEO] Complex on the Transdermal Ability of Clactolide Topical Preparations

[0178] 1. Preparation of Clarithromycin Gel

[0179] Prescription 1: Clactolide Regular Gel

[0180] prescription

[0181] Process: Weigh propylene glycol and isopropyl alcohol and mix them; add weighed clacol ketone while stirring, and continue stirring for 20 minutes; add purified water and sieved high-viscosity hydroxypropyl cellulose HPC-H, and continue stirring until the mixture is uniform; use 10% citric acid aqueous solution to adjust the pH to 4.2-4.9; fill into aluminum tubes for storage.

[0182] Prescription 2: Clactolide-complex gel

[0183] prescription:

[0184] Process: Weigh propylene glycol and isopropyl alcohol and mix them; add weighed clacodone and [OLO] [MEO] while stirring, and continue stirring for 20 minutes; add purified water and sieved HPC-H, and continue stirring until the mixture is uniform; adjust the pH to 4.2-4.9 with 10% citric acid aqueous solution; fill into aluminum tubes for storage.

[0185] Prescription 3: Clactolide gel (ligand added separately)

[0186] prescription:

[0187] Process: Weigh propylene glycol, isopropyl alcohol, and oleyl alcohol and stir; add weighed clacodone and menthol (the molar ratio of menthol to oleyl alcohol is 1:1) while stirring, and continue stirring for 20 minutes; add purified water and sieved HPC-H and continue stirring until the mixture is uniform; adjust the pH to 4.2-4.9 with 10% citric acid aqueous solution; fill into aluminum tubes for storage.

[0188] 2. In vitro transdermal test (IVPT) investigation

[0189] IVPT test method

[0190] The IVPT test was conducted on ex vivo skin from Bama miniature pigs (aged ≤ 3 months) using a Franz diffusion cell. The receiving fluid was collected 24 hours later. After the test, the skin was cleaned and then the tissue was broken down to obtain an intradermal retained extract. The obtained sample was tested using HPLC to determine the clacodone content in the sample.

[0191] IVPT test results

[0192] Table 2 IVPT test results of prescriptions 1 to 3

[0193] The IVPT results are shown in Table 2 and Figure 13. The results show that Formulation 1, a standard clacodone gel, had a transdermal penetration rate comparable to that of the control formulation, Winlevi. When oleyl alcohol and menthol were added as monomers to the formulation (Formulation 3), transdermal permeation increased, but intradermal retention remained unchanged. However, when a complex of oleyl alcohol and menthol was prepared and added to the formulation (Formulation 2), both transdermal permeation and intradermal retention increased significantly. This indicates that oleyl alcohol and menthol have a relatively weak permeation-enhancing effect when used as monomers, but their permeation-enhancing effect is significantly enhanced when they form a complex.

[0194] Example 6 Effects of Other Compounds on the Transdermal Ability of Clactolide External Formulations

[0195] 1. Preparation of Clarithromycin Gel Containing Other Compounds

[0196] Prescription 4: Clactolide-Oleyl Alcohol / Lauryl 2 [OLO] [LRA] Gel

[0197] prescription:

[0198] Process: Weigh propylene glycol and propanol and mix them; add weighed clacodone and 2[OLO][LRA] while stirring, and continue stirring for 20 minutes; add purified water and sieved HPC-H and continue stirring until the mixture is uniform; adjust the pH to 4.2-4.9 with 10% citric acid aqueous solution; fill into aluminum tubes for storage.

[0199] Prescription 5: Clactolide-Oleyl Alcohol / Malic Acid 4 [OLO] [MLA] Gel

[0200] prescription:

[0201] Process: Weigh propylene glycol and isopropyl alcohol and stir; add weighed clacodone and 4[OLO][MLA] while stirring, and continue stirring for 20 minutes; add purified water and sieved HPC-H, and continue stirring until the mixture is uniform; adjust the pH to 4.2-4.9 with 10% citric acid aqueous solution; fill into aluminum tubes for storage.

[0202] 2. In vitro transdermal test (IVPT) investigation

[0203] IVPT test method

[0204] The IVPT test was conducted on ex vivo skin from Bama miniature pigs (aged ≤ 3 months) using a Franz diffusion cell. The receiving fluid was collected 24 hours later. After the test, the skin was cleaned and then the tissue was broken down to obtain an intradermal retained extract. The obtained sample was tested using HPLC to determine the clacodone content in the sample.

[0205] IVPT test results

[0206] Table 3 IVPT test results of prescriptions 1, 4, and 5

[0207] The IVPT test results are shown in Table 3 and Figure 14. The results demonstrate that, compared to conventional gels, the addition of 2[OLO][LRA] or 4[OLO][MLA] increases the intradermal retention of clacodone. Furthermore, these two complexes have a higher affinity for clacodone, making it less susceptible to transdermal absorption and allowing it to exert its effects more effectively within the dermis, thus demonstrating a certain degree of targeting.

[0208] The above describes the embodiments of the present invention. However, the present invention is not limited to the above embodiments. Any modifications, equivalent replacements, improvements, etc. made within the spirit and principles of the present invention shall be included in the scope of protection of the present invention.

Claims

1. A complex consisting of [A] and [B], or consisting of [A], [B] and [C]; in, [A] is selected from long-chain fatty alcohols or polyols; [B] and [C] are the same or different and are independently selected from terpene alcohols or organic acids.

2. The composite according to claim 1, characterized in that Long-chain fatty alcohols are C6-30 long-chain fatty alcohols; Preferably, the long-chain fatty alcohols include long-chain saturated fatty alcohols and long-chain unsaturated fatty alcohols; Preferably, the long-chain fatty alcohols are selected from, for example, decyl alcohol, undecanol, lauryl alcohol, tridecanol, myristyl alcohol, pentadecyl alcohol, palmityl alcohol, heptadecanol, stearyl alcohol, oleyl alcohol, and linoleyl alcohol; Preferably, the polyols are selected from polyols containing two or more hydroxyl groups; Preferably, the polyols are selected from C2-20 polyols; Preferably, the polyols are selected from glycerol, propylene glycol, 1,3-butanediol, 1,2-pentanediol, for example; Preferably, the terpene alcohols are terpene derivatives containing alcoholic hydroxyl groups; Preferably, the terpene alcohols are selected from menthol, linalool, eucalyptol, santalol, nerol, geraniol, citronellol, terpineol, retinol, and ambroxol; Preferably, the organic acid is selected from citric acid, citric acid, malic acid, lauric acid, geranic acid, and lactic acid.

3. The composite according to claim 1 or 2, characterized in that The molar ratio of [A] to [B] is 1:10-10:1, preferably 1:5-5:1; Preferably, the molar ratio of [A], [B], and [C] is 1:10:10-10:1:1, preferably 1:5:5-5:1:1; Preferably, the complex is selected from any one of the following: a complex composed of oleyl alcohol and menthol, a complex composed of oleyl alcohol and citric acid, a complex composed of oleyl alcohol and lauric acid, a complex composed of oleyl alcohol and malic acid, a complex composed of oleyl alcohol and linalool, a complex composed of glycerol and malic acid, and a complex composed of myristyl alcohol, menthol and linalool; preferably, the complex is selected from any one of the following: oleyl alcohol / menthol [OLO][MEO], oleyl alcohol / citric acid [OLO][CA], oleyl alcohol / lauric acid 2[OLO][LRA], oleyl alcohol / malic acid 4[OLO][MLA], oleyl alcohol / linalool [OLO][LNO], glycerol / malic acid 2[GLY]3[MLA], myristyl alcohol / menthol / linalool [MRO][MEO][LNO].

4. A method for preparing the composite according to any one of claims 1 to 3, wherein the method is as follows: The composite is obtained by stirring [A] and [B] while keeping warm, or stirring [A], [B] and [C] while keeping warm.

5. Use of the complex according to any one of claims 1 to 3 in improving drug stability; Preferably, one end of the drug molecular structure is a hydroxyl group and the other end is an alkyl ester group; Preferably, the chemical structure of the drug is as shown in formula (I): in, M is selected from a drug nucleus group; L1 is selected from a bond, -C1-4 alkylene-carbonyl-, C1-4 alkylene-; R1 is selected from C1-10 alkyl.

6. Use of the complex according to any one of claims 1 to 3 in inhibiting transesterification reactions within drug molecules; Preferably, one end of the drug molecular structure is a hydroxyl group and the other end is an alkyl ester group; Preferably, the chemical structure of the drug is as shown in formula (I): in, M is selected from a drug nucleus group; L1 is selected from a bond, -C1-4 alkylene-carbonyl-, C1-4 alkylene-; R1 is selected from C1-10 alkyl.

7. Use of the complex according to any one of claims 1 to 3, wherein the complex is used to enhance the efficacy of a drug; Preferably, the drug is selected from steroid drugs; for example, clacoterone; Preferably, the drug effect is selected from: antagonism of hormones, oil-inhibiting effect on cortical cells, anti-inflammatory effect, etc.

8. A composition or preparation comprising the complex according to any one of claims 1 to 3 and a drug; Preferably, the drug is selected from steroid drugs; for example, clacoterone; Preferably, in the composition or preparation, the mass ratio of the drug to the above-mentioned complex is 0.1%-20%:1%-30%.

9. A method for preparing the composition or preparation according to claim 8, wherein the method is as follows: The components are stirred and mixed to obtain the composition or preparation.

10. Use of the composition or preparation according to claim 8 in the preparation of a medicament for treating acne, alopecia, and genital warts.

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