Cyclopeptide glass and pharmaceutical composition glass containing cyclopeptide

Abstract

The present invention discloses a cyclopeptide glass and a pharmaceutical composition glass containing a cyclopeptide. The cyclopeptide glass of the present invention can exert both the effect of a drug and the function of a drug adjuvant, and compared with crystals and conventional pharmaceutical dosage forms and adjuvants, it can effectively accelerate the dissolution rate of a drug and improve the bioavailability of a drug, and is widely applied in the fields of drug delivery and sustained release, such as antitumor, antiviral, antibacterial, blood sugar control, immunoregulation, and neuroregulation.

Description

[Technical field]

[0001] The present invention belongs to the technical field of medicine, and relates to a new dosage form and a new adjuvant for cyclopeptide drugs, in particular to cyclopeptide glass and cyclopeptide pharmaceutical composition glass. It also relates to a manufacturing method for converting poorly soluble cyclopeptide drugs into glass by effectively utilizing the related technology of poorly soluble drugs to form glass drugs. This cyclopeptide glass is useful in the drug delivery field such as antitumor, antiviral / antibacterial, blood sugar control, immunoregulation, and neuroregulation, especially in sustained drug release therapy. [Background technology]

[0002] Cyclopeptides are condensed cyclized amino acids by peptide bonds, and their structure has a certain conformational constraint effect. Cyclopeptides are extremely stable against chemical, thermal and biodegradation due to their specific topological structure. Cyclopeptides have a large number of hydrogen bond acceptors and donors, and hydrogen bonds are one of the main ways of drug-receptor interaction, so cyclopeptides often have certain biological and pharmacological activities, making them important pharmacophores in medicinal chemistry. In addition, cyclopeptides, due to their restricted conformation, show better specificity and target affinity than linear peptides, and therefore show a wide range of biological and pharmacological activities, such as antitumor, antiviral, antibacterial, antiaging, immunomodulation, memory enhancement, and blood glucose regulation.

[0003] However, due to the remarkable rigid structure and very strong hydrogen bond formation ability, cyclopeptide molecules have the defects of poor water solubility, easy crystallization, and low bioavailability, which seriously affect their medicinal activity. At present, researchers at home and abroad have adopted a variety of formulation methods to improve the above-mentioned shortcomings, including several strategies such as direct chemical modification of cyclopeptides (e.g., N-methylation modification) or encapsulation in nanocarriers (e.g., lipids and micelles). However, problems such as complicated synthesis processes, potential toxicity and low loading of cyclopeptides are still unavoidable.

[0004] Glassy is a metastable amorphous structure that retains the biological and pharmacological activity of cyclopeptide molecules compared to the more thermodynamically stable crystals. Glassy cyclopeptide drugs exhibit higher surface free energy and higher dispersity compared to crystalline cyclopeptide drugs, which can effectively improve the dissolution rate and bioavailability of cyclopeptides.

[0005] In particular, the cyclopeptide glass of the present invention can replace conventional pharmaceutical adjuvants and achieve functions such as dispersion, solubilization, adhesion, controlled release, etc. In particular, it can be used as a substitute for castor oil pharmaceutical adjuvants to effectively avoid severe allergic reactions caused by histamine.

[0006] There have been no published reports on cyclopeptide glasses, especially on glasses based on pharmaceutical compositions with biological / pharmacological activity or sustained-release functions, and on methods for converting poorly soluble cyclopeptides into glass-like forms.

[0007] Surprisingly, the present invention finds that cyclopeptides and their derivatives can be processed into cyclopeptide glass and cyclopeptide pharmaceutical composition glass by a specific manufacturing process.The present invention also finds a method for converting poorly soluble cyclopeptide drugs into glass, which effectively improves the dissolution rate and bioavailability of cyclopeptide drugs.

[0008] The present invention is based on this discovery. The cyclopeptide glass discovered in this invention is expected to be widely applied as an active drug or pharmaceutical adjuvant in the fields of drug delivery and sustained release, including antitumor, antiviral, antibacterial, blood sugar control, immunomodulation, and neuromodulation. Summary of the Invention [Problem to be solved by the invention]

[0009] The main object of the present invention is to provide a cyclopeptide glass, a cyclopeptide pharmaceutical composition glass, and a method for converting a poorly soluble cyclopeptide into a glass-like state. In the present invention, the glass means an amorphous solid exhibiting a glass transition phenomenon, and the glass-like state means a disordered structure that retains glass-like properties, has no crystalline structure in X-ray diffraction detection, and has a determined glass transition temperature. That is, the present invention provides a method for converting a poorly soluble cyclopeptide from a crystalline state to a glass-like state, which effectively improves the in vitro dissolution rate and solubility of a poorly soluble cyclopeptide, and improves the bioavailability of a poorly soluble cyclopeptide drug. [Means for solving the problem]

[0010] In a first aspect, the present invention provides a glass based on a cyclopeptide, the cyclopeptide being a cyclopeptide represented by structural formula 1 and / or a salt thereof, the cyclopeptide being one type or a combination of two or more types. [ka] (A1 to A n are independently one or a combination of more selected from glycine, alanine, valine, leucine, isoleucine, methionine (Met), proline, tryptophan, serine, tyrosine, cysteine, phenylalanine, asparagine, glutamine, threonine, aspartic acid, glutamic acid, lysine, arginine, histidine, selenocysteine, and pyrrolysine; R1~R n are independently selected from H or other modifiable groups, the modifiable groups being preferably methyl, alkyl, phosphate, acetyl, formyl, fatty acid, benzoyl, amide, ester, 9-fluorenylmethoxycarbonyl, t-butoxycarbonyl; n ≧2, preferably 2≦n≦15; A1 to A n is linked by condensation with amino acids.

[0011] Preferably, the cyclopeptide has biological activity and / or pharmacological activity, more preferably, has antibacterial, antiviral, antitumor, blood glucose regulating, immunoregulating activity, and antibacterial and antiviral cyclopeptides include, but are not limited to, the following structures and similar structures:

[0012] [Table 1-1]

[0013] [Table 1-2]

[0014] Antitumor cyclopeptides include, but are not limited to, the following structures and similar structures:

[0015] [Table 2-1]

[0016] [Table 2-2]

[0017] Immunomodulatory cyclopeptides include, but are not limited to, the following structures and similar structures: [Table 3]

[0018] Blood glucose regulating cyclopeptides include, but are not limited to, the following structures and similar structures: [Table 4]

[0019] Cardiovascular and blood related cyclopeptides include, but are not limited to, the following structures and similar structures: [Table 5]

[0020] Other active cyclopeptides include, but are not limited to, the following structures and similar structures: [Table 6]

[0021] Preferably, the cyclopeptide derivative is a molecule having a skeleton similar to the above-mentioned cyclopeptide molecular structure, an isomer, or a salt thereof.

[0022] Preferably, the cyclopeptide is a cyclopeptide having poor water solubility, and the term "poor water solubility" means that the maximum solubility concentration of the cyclopeptide in pure water at room temperature and pressure is 5 wt % or less.

[0023] In a second embodiment, the glasses are based on cyclopeptides, characterized in that they are made exclusively from cyclopeptides and their salts or that the glasses also contain other pharmaceutical adjuvants and / or pharmaceutical active ingredients.

[0024] The pharmaceutical composition glass comprises one or more components selected from the group consisting of a cyclopeptide or a cyclopeptide derivative, another medicament active ingredient or a pharma- ceutical acceptable salt thereof, and a pharmaceutical adjuvant, and preferably contains at least one active cyclopeptide; The pharmaceutical active ingredient includes one or a mixture of two or more of non-cyclopeptide drug molecules having effects such as antibacterial / antiviral, antitumor, blood sugar regulation, immunoregulation, and antipsychotic. Preferably, the antibacterial / antiviral drug is penicillin, cephalexin, amikacin, norfloxacin, nitrofurantoin, metronidazole, amantadine, acyclovir, zidovudine, or ribavirin, and the antitumor drug is cisplatin, doxorubicin, vincristine, paclitaxel, or docetaxel. , gemcitabine, camptothecin, hydroxycamptothecin, irinotecan, etoposide, dexamethasone, fluorouracil, cyclophosphamide; blood glucose regulators are metformin, repaglinide, nateglinide, insulin; immunomodulators are thymopentin, tripterygium glycosides, triptolide, tacrolimus, interferon, krestin; antipsychotics are chlorpromazine, haloperidol, risperidone, paliperidone, ziprasidone; Pharmaceutical adjuvants include fillers, wetting agents, binders, disintegrants, lubricants and others, preferably fillers are starch, lactose, mannitol, wetting agents / binders are sodium carboxymethylcellulose, hydroxypropylcellulose, disintegrants are sodium carboxymethylstarch, low-substituted hydroxypropylcellulose, cross-linked polyvinylpyrrolidone, croscarmellose sodium, lubricants are polyethylene glycol, magnesium stearate, hydrogenated vegetable oil, others are sodium dodecyl sulfate, The mass ratio of the cyclopeptide is 1 to 100%, preferably 1% to 50%; The mass ratio of other medicamentously active ingredients is 1 to 20%, preferably 1 to 10%; The mass ratio of the pharmaceutical adjuvant is 0-50%, preferably 0-5%, more preferably 0%, that is, the cyclopeptide glass completely replaces the conventional pharmaceutical adjuvant.

[0025] In a third aspect, glasses based on cyclopeptides are characterized in that, as pharmaceutical adjuvants, they can completely or partially replace conventional pharmaceutical adjuvants.

[0026] In a fourth aspect, there is provided a method for producing a cyclopeptide glass drug or a cyclopeptide glass adjuvant by converting a poorly soluble cyclopeptide into a glass form, the method comprising the steps of: (1) controlling the temperature of the raw material during ball milling to 0 to 50°C, preferably 10 to 30°C, and thoroughly ball milling one or more types of poorly soluble cyclopeptides; (2) by the "heating-quenching" method, specifically, by heating the sufficiently pulverized cyclopeptide raw material in an inert gas atmosphere to a temperature close to the melting point temperature, and then by keeping the temperature for a predetermined period of time, the cyclopeptide raw material is transferred to an annealing furnace for annealing to obtain a cyclopeptide glass drug or a cyclopeptide glass adjuvant; The heating temperature is the melting point temperature (T m ) ±50 to 250K, preferably T m 50-100K higher than The incubation time is 0 min to 30 h, preferably 15 to 30 min. The annealing temperature is the glass transition temperature (T g ) ±50 to 150K, preferably T g 50 to 100K lower than The annealing time is 30 min to 2 h, preferably 30 min to 1 h. T m and T g is measured by thermogravimetric analysis and differential scanning calorimetry, and the heating and cooling rates are 2 to 50 Kmin -1 The present invention is characterized in that:

[0027] In a fifth aspect, a method for producing a poorly soluble cyclopeptide pharmaceutical composition glass, comprising the steps of: (1) mixing the poorly soluble cyclopeptide and other medicamentously active ingredients, and then thoroughly milling the raw materials in a ball mill at a temperature of 0 to 50°C, preferably 10 to 30°C; Step (2) similar to step (2) of the fourth aspect; and step (3) of producing a pharmaceutical composition by one or a combination of two or more of the following: tabletting, wet granulation, fluidized bed granulation, coating, spray granulation, program casting, and 3D printing, using both the glass obtained in step (2) and the pharmaceutical adjuvant.

[0028] Alternatively, the following manufacturing method is provided, which mainly comprises: (1) mixing the poorly soluble cyclopeptide and other medicamentously active ingredients, and then thoroughly milling the mixture while controlling the temperature of the raw materials during ball milling to 0 to 50°C, preferably 10 to 30°C; The powder from step (1) is heated in an inert gas atmosphere and kept at the temperature for a predetermined period of time. The temperature is then increased to the melting point temperature (T m ) ±50 to 250K, preferably T m (2) setting the temperature at 50 to 100 K higher than the temperature at which the temperature was set, and setting the temperature retention time to 0 min to 30 h, preferably 15 to 30 min; Step (3) of dissolving other medicamentously active ingredients in a good solvent and a co-solvent, the good solvent being preferably ethanol, hexafluoroisopropanol, methylene chloride, carbon tetrachloride, acetone, ethyl acetate, and the co-solvent being preferably one or a mixture of two or more of sodium benzoate, dimethylacetamide, urea, and Tween; Step (4) of uniformly mixing the medicament active ingredient solution obtained in step (3) and the molten cyclopeptide of step (1); (5) subjecting the mixture obtained in step (4) to the temperature set in step (2) and subjecting the solvent to rotary evaporation under reduced pressure; The mixture obtained in step (5) is transferred to an annealing furnace and annealed at a temperature above the glass transition temperature (T g ) ±50 to 150K, preferably T g (6) setting the temperature at 50 to 100 K lower than the temperature at which the annealing treatment was performed and setting the annealing time to 30 min to 2 h, preferably 30 min to 1 h; and step (7) of producing a pharmaceutical composition using both the glass obtained in step (6) and the pharmaceutical adjuvant by one or a combination of two or more of the following: tabletting, wet granulation, fluidized bed granulation, coating, spray granulation, program casting, and 3D printing.

[0029] Alternatively, the following manufacturing method is provided, which mainly comprises: Step (1) of dissolving the poorly soluble cyclopeptide and other medicamentously active ingredients in a good solvent and a co-solvent, the good solvent being preferably ethanol, hexafluoroisopropanol, methylene chloride, carbon tetrachloride, acetone, or ethyl acetate, and the co-solvent being preferably one or a mixture of two or more of sodium benzoate, dimethylacetamide, urea, and Tween; The mixed solution obtained in step (1) is heated in an inert gas atmosphere, the solvent is evaporated by rotary evaporation under reduced pressure, and the heated temperature is kept at a temperature equal to the melting point temperature (T m ) ±50 to 250K, preferably T m (2) setting the temperature at 50 to 100 K higher than the temperature at which the temperature was set, and setting the temperature retention time to 0 min to 30 h, preferably 15 to 30 min; The mixture obtained in step (2) is transferred to an annealing furnace and annealed at a temperature above the glass transition temperature (T g ) ±50 to 150K, preferably T g (3) setting the temperature at 50 to 100 K lower than the temperature at which the annealing treatment was performed and setting the annealing time to 30 min to 2 h, preferably 30 min to 1 h; and step (4) of producing a pharmaceutical composition by one or a combination of two or more of the following: tabletting, wet granulation, fluidized bed granulation, coating, spray granulation, program casting, and 3D printing, using both the glass obtained in step (3) and the pharmaceutical adjuvant.

[0030] In a sixth aspect, according to the cyclopeptide glass drug, the cyclopeptide glass adjuvant, the cyclopeptide pharmaceutical composition glass and the manufacturing method described in the first to fifth aspects, the glass pharmaceutical dosage form may be an oral agent, a patch, a subcutaneous implant, a stent material, and a microneedle device.

[0031] In a seventh aspect, the pharmaceutical dosage form according to the sixth aspect, wherein the process is one or more of: tabletting, dry granulation, high shear wet granulation, fluid bed granulation, capsule filling, microcapsule embedding, coating, spray drying, spray congealing, photolithography, programmed casting, microneedle array, 3D printing.

[0032] In an eighth aspect, the cyclopeptide glass, and the cyclopeptide pharmaceutical composition glass, and the cyclopeptide drug glass dosage form of the present invention have the following advantages and beneficial effects: (1) It can exert both the effect of a drug and the function of a drug adjuvant. (2) It can effectively accelerate the dissolution rate of drugs, improve the bioavailability of drugs, and reduce the side effects of drugs. (3) It has good biocompatibility and biodegradability. (4) High thermal and chemical stability.

[0033] In a ninth aspect, the cyclopeptide glass, and the cyclopeptide pharmaceutical composition glass, and the cyclopeptide drug glass dosage form of the present invention are useful in the drug delivery fields such as antitumor, antiviral / antibacterial, glycemic control, immunomodulation, neuromodulation, etc., particularly in sustained drug release therapy. Effect of the Invention

[0034] The raw material of the cyclopeptide glass of the present invention includes any one or more combinations of cyclopeptides, particularly cyclopeptides having biological activity and pharmacological activity or their pharma- ceutically acceptable derivatives and salts, which generally have poor water solubility, are prone to crystallization, and have low bioavailability. The present invention proposes a novel method for converting poorly soluble cyclopeptide drugs into glass, including a method for producing poorly soluble cyclopeptides into glass, and a method for producing glass using both poorly soluble cyclopeptides and other active drug components. The present invention provides a method for converting poorly soluble drugs into glass, forming a drug glass dosage form, and realizing poorly soluble active cyclopeptides and sustained drug release. The obtained cyclopeptide glass is highly biocompatible, biodegradable, and has high thermal and chemical stability, and may be manufactured into a synthetic oral agent, a patch, a subcutaneous implant, a stent material, or a microneedle device by techniques such as spray drying, solid dispersion, and layered manufacturing. The obtained cyclopeptide glass can exert the effect of drugs and the function of drug adjuvants. Compared with crystals and traditional pharmaceutical dosage forms or adjuvants, cyclopeptide glass can effectively accelerate the dissolution rate of drugs and improve the bioavailability of drugs, and is widely applied in the fields of drug delivery and sustained release, such as antitumor, antiviral, antibacterial, blood sugar control, immunomodulation, and neuromodulation. [Brief description of the drawings]

[0035] [Figure 1] 1A is a photograph of the CsA glass produced in Example 1 at room temperature, and FIG. 1B is a photograph of the CsA glass obtained by a casting process, demonstrating the processability of cyclopeptide glass. [Diagram 2] This is the nuclear magnetic resonance hydrogen spectrum of the CsA glass produced in Example 1. Compared to the CsA raw material, no significant change was observed in the peaks, demonstrating that the chemical composition of the cyclopeptide raw material molecules does not change even after heat melting and annealing treatments. [Diagram 3]1 is a differential scanning calorimetry (DSC) diagram of the CsA glass prepared in Example 1, which demonstrates the glass-like structure of the CsA glass, and the glass transition temperature of the CsA glass is Tg=100.4K. [Figure 4] 1 is an X-ray diffraction (XRD) pattern of the CsA glass produced in Example 1, which demonstrates that the CsA glass has an amorphous structure. [Diagram 5] 1 shows the stability test results of the CsA glass produced in Example 1, which demonstrates the high stability of the CsA glass. [Figure 6] 1 shows the drug release curve of the CsA glass prepared in Example 1, demonstrating that the drug can be released in a sustained manner. [Figure 7] 4 shows immune-related indexes of mice orally administered with the CsA glass produced in Example 1. [Figure 8] 1 is an XRD spectrum of the pharmaceutical composition glass based on CFP and the tumor chemotherapy drug PTX of Example 2, which has an amorphous structure. [Figure 9] The glass capsule of the pharmaceutical composition prepared in Example 2 was decomposed in artificial gastric juice (prepared according to the method described in the Chinese Pharmacopoeia), and the decomposition property of the glass capsule of the pharmaceutical composition was demonstrated. [Figure 10] 1 shows an in vivo drug-time curve in mice after intragastric administration of the glass capsule pharmaceutical composition prepared in Example 2. [Figure 11] 1 shows a tumor inhibition curve of experimental mice orally administered with the glass capsule of the pharmaceutical composition prepared in Example 2, demonstrating the antitumor effect of the glass capsule of the pharmaceutical composition. [Figure 12] 1 shows a weight change curve of experimental mice orally administered with the glass capsules of the pharmaceutical composition prepared in Example 2, demonstrating high biosecurity. [Figure 13] FIG. 1 shows the changes in organ coefficients of experimental mice orally administered with the glass pharmaceutical composition capsules prepared in Example 2, demonstrating high biosecurity. [Figure 14] 1 shows a melting image and XRD data of the pharmaceutical composition glass of Example 3, demonstrating the conversion from crystal to glass. [Figure 15] 1 shows the biocompatibility test results of the pharmaceutical composition glass prepared in Example 3, demonstrating high biocompatibility. [Figure 16] 1 is a bacterial growth inhibition curve of the pharmaceutical composition glass coating prepared in Example 3. [Figure 17] The inhibitory effect of the pharmaceutical composition glass tablet prepared in Example 3 against Escherichia coli and Staphylococcus aureus. [Figure 18] 1 shows the time-dependent decomposition curve of the glass pharmaceutical composition embedding agent prepared in Example 3 after being embedded subcutaneously. [Figure 19] 1 shows the blood glucose control curve of mice with idiopathic type II diabetes using the glass tablet of the pharmaceutical composition prepared in Example 4, which demonstrates that the blood glucose of the mice can be effectively controlled. [Figure 20] This is a time-dependent change curve of mouse weight when the pharmaceutical composition glass tablet prepared in Example 4 was intragastrically administered to idiopathic type II diabetes mice, demonstrating that the mouse weight can be effectively controlled. [Figure 21] Thirty days after intragastric administration of the glass tablet of the pharmaceutical composition prepared in Example 4 to mice with idiopathic type II diabetes, a high concentration of glucose (2.5 g kg-1) was orally administered. A curve was plotted by measuring the blood glucose concentration in the sole blood of the mice, demonstrating that the mice in the experimental group had higher glucose tolerance. [Figure 22] This is the DSC curve of the CPY-CFP-CLP cyclopeptide mixed glass prepared in Example 5. The glass transition temperatures of the single and mixed glasses were obtained, and it was found that when multiple types of cyclopeptides are mixed, the tendency of the single cyclopeptide to crystallize can be effectively suppressed. [Figure 23] 1 shows the CLP molecule release curves of the cyclopeptide-mixed glass and CLP crystals prepared in Example 5, which indicates that the cyclopeptide-mixed glass accelerates the dissolution rate of CLP-active cyclopeptides. [Figure 24] 1 shows the XRD pattern of the pharmaceutical composition glass prepared in Example 5, the pharmaceutical composition glass being amorphous. [Diagram 25] This is statistical data on spontaneous activity ability and anxiety-like behavior of mice in an open field experiment. As a result, compared with normal mice, early social isolation mice showed a clear decrease in the activity distance in the central area, but after treatment with glass microneedle patches, a significant improvement was observed. [Figure 26] The number of times the mice entered the open and closed arms and the time spent in the two arms within 5 minutes in a maze experiment were measured. Compared to normal mice, the early social isolation mice showed a significant decrease in the number of times they entered the open arms and a shorter time spent in the open arms, but after treatment with the glass microneedle patch, a significant improvement was observed. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0036] The technical solutions of the present invention are described in detail below through examples, but the scope of the invention is not limited to the following.

[0037] Example 1 The method for preparing cyclopeptide glasses based on cyclosporin A (CsA) includes the following steps. (1) The CsA powder was placed in a ball mill and crushed uniformly. The ball mill was kept at 25°C and then transferred to a crucible. (2) The crucible containing the CsA powder in step (1) was placed in a heating apparatus in a N2 atmosphere. (3) Heat the device of step (2), i.e., 10Kmin -1 The crucible was heated from room temperature to 573.15 K at a heating rate of 0.15° C. and kept at this temperature for 20 min. (4) The temperature of the device in step (3) is lowered, i.e., 10Kmin -1 The temperature of the crucible was lowered to 273.15 K at a rate of 0.15° C., and the glass was annealed by keeping the temperature in the annealing furnace for 30 minutes to obtain a CsA glass.

[0038] FIG. 1 shows (a) an actual image of the CsA glass produced in Example 1 at room temperature, and (b) an actual image of the CsA glass obtained by a casting process, demonstrating the processability of the cyclopeptide glass.

[0039] FIG. 2 shows the nuclear magnetic resonance hydrogen spectrum of the CsA glass produced in Example 1. No obvious change was observed in the peaks compared to the CsA raw material, revealing that the chemical composition of the cyclopeptide raw material molecules did not change even after undergoing heating, melting and annealing treatments.

[0040] FIG. 3 is a DSC diagram of the CsA glass produced in Example 1, which demonstrates the glassy structure of the CsA glass. The glass transition temperature of the CsA glass is T g =100.4K.

[0041] FIG. 4 is an X-ray diffraction (XRD) pattern of the CsA glass produced in Example 1, which demonstrates that the CsA glass has an amorphous structure.

[0042] FIG. 5 shows the stability test results of the CsA glass produced in Example 1, which demonstrates the high stability of the CsA glass.

[0043] Figure 6 shows the drug release curve of the CsA glass prepared in Example 1, which demonstrates that the drug can be released in a sustained manner. Specifically, the initial mass of the CsA glass was 20±1 mg, the sustained release solvent was 10 mL of phosphate buffer (0.01 M, pH=6.8), and the stirring speed was 100 rpm. -1 The eluted CsA is detected using a dissolution tester and high performance liquid chromatography, and the drug release curve is plotted.

[0044] Figure 7 shows immune-related indicators of mice orally administered the CsA glass prepared in Example 1. Specifically, healthy male Kunming mice (body weight 20±1 g, 4-5 weeks old) were randomly divided into two groups of 8 mice each and fed a normal diet. The mice in the experimental group were administered 40 mg / kg -1The mice in the control group were intragastrically administered with a dose of 100 mg / kg / day, 5 times in total, every other day, and the mice in the control group were intragastrically administered with the same mass of saline every other day. The mice were euthanized the day after the last oral administration, and the corresponding indexes were detected. The mice were dissected, and the thymus and spleen were removed, the surface blood was washed off with phosphate-buffered saline (PBS), and the moisture was absorbed with filter paper, after which they were weighed on an electronic balance, and the organ weight index was calculated by dividing the organ weight (mg) by the total weight of the mouse (g). Peripheral blood was collected from the mice, and CD3 + (PE-Cy5) monoclonal antibody, CD4 + (PE) Monoclonal antibody, CD8 + The cells were mixed with (APC) monoclonal antibody, shaken uniformly, incubated in the dark, and after cell lysis, centrifuged and analyzed by flow cytometer.

[0045] Example 2 The method for preparing a pharmaceutical composition glass based on phenylalanine-proline cyclodipeptide (CFP) and paclitaxel (PTX) includes the following steps: (1) The CFP powder was placed in a ball mill, pulverized, and mixed uniformly. The ball mill was kept at 37°C and then transferred to a container. (2) Heat the vessel of step (1), i.e., 20Kmin -1 The container was heated from room temperature to 473.15 K at a heating rate of 0.01°C and kept at this temperature for 30 min. (3) The temperature of the container in step (2) is lowered, i.e., 10Kmin -1 The temperature of the vessel was decreased to 333.15 K at a rate of 0.05°C. (4) The PTX powder was placed in a ball mill and ground uniformly. The ball mill was kept at 37°C, and then ethanol was added to completely dissolve the powder, making the molar ratio of CFP to PTX 25:1. (5) The solution from step (4) was added dropwise to the container from step (3), and the container was placed in a reduced pressure rotary evaporator, maintaining the temperature at 333.15 K, to remove the solvent. (6) The temperature of the device in step (5) is lowered, i.e., 10Kmin -1The container was cooled to 315.15K at a cooling rate of 0.01 to 0.15K, and then kept in an annealing furnace for 30min to obtain a pharmaceutical composition glass based on CFP and a tumor chemotherapy drug PTX. The glass was completely dissolved in an organic solution of HFIP, and the concentration of PTX was quantified by high performance liquid chromatography. The calculated amount of PTX was 3.5±0.2%. (7) Cyclopeptide glass oral dosage forms were prepared by a manufacturing process of spray drying and capsule filling.

[0046] FIG. 8 is the XRD spectrum of the pharmaceutical composition glass based on CFP and the tumor chemotherapy drug PTX of Example 2, which demonstrates that the pharmaceutical composition of the present invention has an amorphous structure.

[0047] Figure 9 shows the decomposition of the pharmaceutical composition glass capsule prepared in Example 2 in artificial gastric fluid (according to the preparation method described in the Chinese Pharmacopoeia), which demonstrates the decomposability of the pharmaceutical composition glass capsule. The preparation steps of the pharmaceutical composition glass capsule are to crush the pharmaceutical composition glass into powder by a homogenizer, and then fill the above powder into a gelatin capsule shell.

[0048] Figure 10 shows the in vivo drug-time curve of mice intragastrically administered the pharmaceutical composition glass capsules prepared in Example 2. Specifically, mice were administered 8 mg kg -1 After intragastric administration at a dose of 100 mg / kg / day, peripheral blood was collected from the mice by collecting blood from the orbit over time, cells were lysed and eluted with a solvent, and the filtered solution was detected by high performance liquid chromatography to calculate the PTX content in the blood and plot the drug-time curve.

[0049] Fig. 11 shows the tumor inhibition curve of experimental mice orally administered with the pharmaceutical composition glass capsule prepared in Example 2, demonstrating the antitumor effect of the pharmaceutical composition glass capsule. Specifically, healthy BALB / c nude mice (body weight 15±1 g, 4-6 weeks old) were randomly divided into two groups of 8 mice each, and 4T1 cells were inoculated subcutaneously into the axilla to establish a 4T1 mouse breast cancer transplantation tumor model. The tumors in the mice were 80±10 mm 3After the mice had grown to a volume of 100 mg kg 2, they were treated with the drug. -1 The mice in the control group were intragastrically administered with the same mass of PBS every other day. The changes in the tumor volume of the mice were measured daily using a caliper and recorded. The tumor volume of the mice was calculated using the formula V=ab 2 / 2, where a is the length of the long side of the tumor and b is the length of the short side. FIG. 12 shows the body weight change curve of experimental mice orally administered with the glass capsules of the pharmaceutical composition prepared in Example 2, demonstrating high biosecurity. 13 shows the change in organ index of experimental mice orally administered with the pharmaceutical composition glass capsules prepared in Example 2, demonstrating high biosecurity. Specifically, the vital organs of the mice (including the heart, liver, spleen, lungs, and kidneys) were dissected on the 35th day and obtained, the surface blood was washed away with PBS, the moisture was absorbed with filter paper, and the organs were weighed on an electronic balance, and the organ weights (mg) were divided by the total weight (g) of the mice to calculate the organ index.

[0050] Example 3 The method for preparing pharmaceutical composition glass based on vancomycin, gramicidin S, and penicillin includes the following steps. (1) Vancomycin, gramicidin S, and penicillin were placed in a ball mill, pulverized, and mixed uniformly. The ball mill was maintained at 37°C and then transferred to a container such that the mass ratio of these three was 2:2:1. (2) A mixed organic solvent of hexafluoroisopropanol and ethanol was added to the mixed powder of step (1) and stirred until completely dissolved. (3) Place the mixed solution obtained in step (2) in a vacuum rotary evaporator and evaporate for 50 Kmin. -1 The temperature was raised from room temperature to 443.15 K at a heating rate of 100° C., and the mixture was kept at this temperature for 30 minutes to remove the organic solvent. (4) The temperature of the device in step (3) is lowered, i.e., 10Kmin -1 The temperature was then lowered to 243.15 K at a rate of 0.05°C and maintained at this temperature for 20 min. (5) To the product obtained in step (4), a pharmaceutical adjuvant was added, comprising 1.5 parts starch, 1 part sodium carboxymethylcellulose, 2.5 parts polyethylene glycol and 6.5 parts hydrogenated vegetable oil, in a total amount of 92.5% by weight. (6) The pharmaceutical composition obtained in step (5) was homogenized, and the glass pharmaceutical composition embedding agent was prepared by a combined technique of tableting and vacuum spray drying.

[0051] FIG. 14 shows a melting image and XRD data of the pharmaceutical composition glass of Example 3, which demonstrates the conversion from crystal to glass.

[0052] Figure 15 shows the biocompatibility test results of the pharmaceutical composition glass prepared in Example 3, which demonstrated high biocompatibility. Specifically, the above glass was processed into a cell culture dish coating with a diameter of 60 mm and a thickness of 0.2 mm using a coater, and used for cell culture (fibroblast 3T3). After 48 h of incubation, the cell viability was tested using the MTT method.

[0053] Figure 16 shows the bacterial growth inhibition curve of the pharmaceutical composition glass coating prepared in Example 3. Specifically, the pharmaceutical composition glass was processed into a tablet of 1 cm x 1 cm x 0.1 cm, and pasted in the center of a bacterial culture dish. The total drug concentration was calculated to be 10 μg / dish. After E. coli and Staphylococcus aureus were mixed uniformly in liquid medium, the mixture was dropped onto the bacterial culture dish, and incubated at 37°C with shaking. The bacterial concentration in the bacterial culture dish was measured over time by the OD value method.

[0054] 17 shows the inhibitory effect of the pharmaceutical composition glass tablet prepared in Example 3 against E. coli and Staphylococcus aureus. Specifically, the pharmaceutical composition glass was processed into a tablet with a diameter of 5 mm and a thickness of 0.2 mm, an Oxford cup (diameter 6 mm) was placed on an agar culture dish, and the tablet was placed in the Oxford cup and adhered to the agar. E. coli and Staphylococcus aureus were mixed uniformly with PBS, then applied to the inside of the culture dish, and cultured in an incubator at 37°C. After 24 hours, the presence or absence of an inhibition circle was observed to determine the inhibitory effect of the pharmaceutical composition glass.

[0055] Figure 18 shows the time-dependent decomposition curve of the pharmaceutical composition glass embedding agent prepared in Example 3 after subcutaneous embedding. Specifically, healthy male Kunming mice (body weight 20±1g, 4-5 weeks old) were randomly divided into two groups of 15 mice each. A 1cm window was surgically opened on the back of the mice in the experimental group, and 30±2.5g of the pharmaceutical composition glass was embedded subcutaneously and sutured. The mice were sampled over time, the pharmaceutical composition glass was removed, the remaining mass was weighed, and the decomposition curve was plotted.

[0056] Figure 19 shows the blood glucose control curve of idiopathic type II diabetes mice using the pharmaceutical composition glass tablet prepared in Example 4, demonstrating that the blood glucose of the mice can be effectively controlled. Specifically, purchased idiopathic type II diabetes model mice (male, 8 weeks old) were divided into two groups of six mice each. The mice in the experimental group were administered 4 mg kg -1 The pharmaceutical composition was administered intragastrically every day at a dose of 100 mg / kg / day, and the control group was administered intragastrically with the same amount of saline, with the test period being 30 days. The mice were sampled daily from their tails and their fasting blood glucose levels were measured using a blood glucose tester. A curve was plotted with the horizontal axis representing time and the vertical axis representing the fasting blood glucose levels of the experimental mice.

[0057] FIG. 20 shows a time course curve of the body weight of idiopathic type II diabetes mice when the glass tablet of the pharmaceutical composition prepared in Example 4 was intragastrically administered to the mice, demonstrating that the body weight of the mice can be effectively controlled.

[0058] FIG. 21 shows the glucose concentration of 2.5 g kg 30 days after intragastric administration of the glass tablet of the pharmaceutical composition prepared in Example 4 to spontaneous type II diabetes mice. -1 This is the tolerance when high-concentration glucose was orally administered, and a curve was plotted by measuring the blood glucose concentration in the plantar blood of the mice, demonstrating that the experimental group mice had higher glucose tolerance.

[0059] FIG. 22 shows the DSC curve of the CPY-CFP-CLP cyclopeptide mixed glass prepared in Example 5. The glass transition temperatures of the single and mixed glasses were obtained, and it was found that when multiple types of cyclopeptides are mixed, the tendency of the single cyclopeptide to crystallize can be effectively suppressed.

[0060] Figure 23 shows the CLP molecule release curves of the cyclopeptide mixed glass and CLP crystals prepared in Example 5. It was found that the cyclopeptide mixed glass accelerated the dissolution rate of the CLP active cyclopeptide. Specifically, the initial mass of the cyclopeptide mixed glass was 15±1 mg, the sustained release solvent was 10 mL PBS (0.01 M, pH=6.8), and the stirring speed was 100 rpm. -1 The eluted CLP was detected using a dissolution tester and high performance liquid chromatograph, and the release curve of the cyclopeptide mixed glass was plotted. As a control, the same mass of CLP crystals was placed in the above-mentioned release solution, stirred and incubated, and the concentration of CLP in the release solution was detected.

[0061] FIG. 24 is an XRD pattern of the pharmaceutical composition glass prepared in Example 5, and the pharmaceutical composition glass is amorphous.

[0062] FIG. 25 shows statistical data on spontaneous activity and anxiety-like behavior of mice when the pharmaceutical glass composition prepared in Example 6 was used in a small animal open field experiment. As a result, compared with normal mice, early social isolation mice showed a clear decrease in the activity distance in the central area, but after treatment with glass microneedle patches, a significant improvement was observed. Specifically, C57BL / 6 early social isolation mice (8 weeks old, male) were divided into two groups, each with an average of 6 mice. One group was not treated specially, and the other group of mice had their backs depilated and then had glass microneedle patches attached. The C57BL / 6 group was a blank control group, containing 6 normal mice. According to the standard, a black open field apparatus (45 cm x 5 cm x 45 cm) was prepared, and the mice were placed in the open field, and the spontaneous activity behavior of the mice was tracked and recorded. The ordinate is the statistical value of the activity distance in the central area when the mice moved in the open field for 1 h.

[0063] FIG. 26 shows the number of mouse entries into the open and closed arms and the time spent in the two arms within 5 minutes when the pharmaceutical composition glass produced in Example 6 was used in a mouse maze experiment. As a result, compared with normal mice, the early social isolation mice had a significantly reduced number of entries into the open arms and a shorter time spent in the open arms, but after treatment with the glass microneedle patch, a significant improvement was observed. Specifically, according to the standard, a maze device was prepared with an arm width of 5 cm, an arm length of 35 cm, a closed arm height of 15 cm, a central platform of 5 cm x 5 cm, and a maze height of about 40 to 55 cm from the ground. The mouse was placed in the center of the platform, and the number of mouse entries into the open and closed arms and the time spent in the two arms were recorded to evaluate the anxiety index.

[0064] Example 4 The method for preparing the pharmaceutical composition glass based on histidine-proline cyclodipeptide and secretide includes the following steps. (1) Histidine-proline cyclodipeptide and secretide were placed in a ball mill, pulverized, and mixed uniformly. The ball mill was kept at 25° C., and then transferred to a container, so that the mass ratio of the two was 10:1. (2) Dichloromethane solvent was added to the mixed powder of step (1) and stirred until completely dissolved. (3) Place the mixed solution obtained in step (2) in a vacuum rotary evaporator and evaporate for 10 km -1 The temperature was raised from room temperature to 535.15 K at a heating rate of 100° C., and the mixture was kept at this temperature for 30 minutes to remove the organic solvent. (4) The temperature of the device in step (3) is lowered, i.e., 10Kmin -1 The temperature was lowered to 283.15 K at a rate of 0.15° C. and the mixture was kept at this temperature for 1 hour to obtain a pharmaceutical composition glass based on histidine-proline cyclodipeptide and secretide. (5) The pharmaceutical composition glass obtained in step (4) was homogenized, and then tablets were prepared by tableting and program casting, and used for oral administration and blood sugar control.

[0065] Example 5 The method for preparing mixed glasses based on proline-tyrosine cyclodipeptide (CPY), phenylalanine-proline cyclodipeptide (CFP), and leucine-proline cyclodipeptide (CLP) includes the following steps: (1) CPY, CFP, and CLP powders were weighed in equal molar ratios and dissolved in hexafluoroisopropanol to form a single cyclopeptide solution. (2) Place the mixed solution obtained in step (1) in a vacuum rotary evaporator and evaporate for 10 km -1 The temperature was raised from room temperature to 473.15 K at a heating rate of 0.015° C., and the mixture was kept at this temperature for 30 minutes to remove the organic solvent. (3) The temperature is lowered in the apparatus of step (2), i.e., 10Kmin -1 The temperature was then lowered to 243.15 K at a rate of 0.15° C. and kept at this temperature for 30 min to obtain a CPY-CFP-CLP cyclopeptide mixed glass.

[0066] Example 6 The method for preparing a pharmaceutical composition glass based on tryptophan-tryptophan cyclodipeptide (CWW) and risperidone includes the following steps. (1) CWW and risperidone powder were weighed in an equal molar ratio and dissolved in hexafluoroisopropanol and methanol, respectively, to form monodisperse solutions. (2) Place the mixed solution obtained in step (1) in a vacuum rotary evaporator and evaporate for 10 km -1 The temperature was raised from room temperature to 573.15 K at a heating rate of 0.015° C., and the mixture was kept at this temperature for 30 minutes to remove the organic solvent. (3) The temperature of the device in step (2) is lowered, i.e., 50Kmin -1 The temperature was lowered to 283.15 K at a rate of 0.15° C. and the mixture was kept at this temperature for 30 minutes to obtain a glass pharmaceutical composition based on CWW and paliperidone. (4) The glass pharmaceutical composition obtained in step (3) was processed into a microneedle patch with a size of 0.8 × 0.8 cm by 3D printing and casting processes, and used for drug transdermal delivery. A C57BL / 6 early social isolation mouse model was established, and a risperidone dose of 5 mg kg was administered. -1 Mice were treated with a subcutaneous microneedle patch of the drug and assessed for schizophrenia-like behavior 14 days later.

Claims

1. 1. Cyclopeptide-based glasses, comprising: The cyclopeptide is a cyclopeptide represented by formula 1 and a salt thereof, and may be one or a combination of two or more thereof; The cyclopeptide-based glass is characterized in that the cyclopeptide preferably has biological and / or pharmacological activity, more preferably antibacterial, antiviral, antitumor, blood glucose regulating and / or immunoregulating activity. 【Chemistry 1】 (However, A １ ~A ｎ are independently selected from glycine, alanine, valine, leucine, isoleucine, methionine (Met), proline, tryptophan, serine, tyrosine, cysteine, phenylalanine, asparagine, glutamine, threonine, aspartic acid, glutamic acid, lysine, arginine, histidine, selenocysteine, and pyrrolysine; R １ ~R ｎ are independently selected from H or other modifiable groups, and the modifiable groups are independently selected from methyl, alkyl, phosphate, acetyl, formyl, fatty acid, benzoyl, amide, ester, 9-fluorenylmethoxycarbonyl, t-butoxycarbonyl; n ≧2, preferably 2≦n≦15; A １ ~A ｎ is linked by condensation with an amino acid.)

2. The antibacterial and antiviral cyclopeptide is one or a combination of multiple types selected from the following cyclopeptides: 【Table 1-1】 【Table 1-2】 The antitumor cyclopeptide is one or a combination of multiple types selected from the following cyclopeptides: 【Table 2-1】 【Table 2-2】 The immunomodulatory cyclopeptide is one or a combination of multiple types selected from the following cyclopeptides: 【Table 3】 The blood glucose-regulating cyclopeptide is one or a combination of multiple types selected from the following cyclopeptides: 【Table 4】 The cardiovascular and blood-related cyclopeptides are one or a combination of multiple cyclopeptides selected from the following: 【Table 5】 The other active cyclopeptides are one or a combination of two or more selected from the following cyclopeptides: 【Table 6】 A glass based on the cyclopeptide of claim 1.

3. The glass based on a cyclopeptide according to claim 1, characterized in that the cyclopeptide is a poorly water-soluble cyclopeptide, and also includes a water-soluble cyclopeptide derivative in which the peptide chain backbone of the poorly water-soluble cyclopeptide is modified.

4. A pharmaceutical composition comprising the glass according to any one of claims 1 to 3, The pharmaceutical composition is in glass form and is characterized in that it is made up of the cyclopeptide alone or further contains a pharmaceutical adjuvant and / or a pharmaceutical active ingredient.

5. A method for producing the cyclopeptide glass according to any one of claims 1 to 3, comprising the steps of: (1) controlling the temperature of the raw material during ball milling to 0 to 50°C, preferably 10 to 30°C, and ball milling one or more cyclopeptides; (2) producing cyclopeptide glass by a "heating-quenching" method, heating the crushed cyclopeptide raw material to near its melting point in an inert gas atmosphere, keeping it at that temperature for a predetermined period of time, and then transferring it to an annealing furnace for annealing.

6. The heating temperature is the melting point temperature (T ｍ ) ±50 to 250K, preferably T ｍ 50 to 100K higher than The incubation time is 0 min to 30 h, preferably 15 to 30 min. The annealing temperature is the glass transition temperature (T ｇ ) ±50 to 150K, preferably T ｇ 50 to 100K lower than The annealing time is 30 min to 2 h, preferably 30 min to 1 h. T ｍ and T ｇ is measured by thermogravimetric analysis and differential scanning calorimetry, and the heating and cooling rates are 2 to 50 Kmin. －１ 6. The method of claim 5, wherein:

7. A method for producing the pharmaceutical composition of claim 4, comprising the steps of: (1) mixing the cyclopeptide with other medicamentously active ingredients and ball milling the mixture while controlling the temperature of the raw materials during ball milling to 0-50°C, preferably 10-30°C; (2) producing cyclopeptide glass by the "heating-quenching" method, heating the crushed cyclopeptide raw material to a temperature close to the melting point temperature in an inert gas atmosphere, and then transferring it to an annealing furnace for annealing to obtain glass; and optionally, a step (3) of preparing a pharmaceutical composition using both the glass obtained in step (2) and a pharmaceutical adjuvant.

8. A method for producing the pharmaceutical composition of claim 4, comprising the steps of: (1) controlling the temperature of the raw material during ball milling to 0 to 50°C, preferably 10 to 30°C, and ball milling one or more cyclopeptides; The powder from step (1) is heated in an inert gas atmosphere and treated by keeping the temperature for a predetermined period of time. The temperature is then increased to the melting point temperature (T ｍ ) ±50 to 250 K, preferably T ｍ (2) setting the temperature at 50 to 100 K higher than the temperature at which the temperature is maintained, and setting the temperature maintenance time at 0 min to 30 h, preferably 15 to 30 min; Step (3) of completely dissolving other medicamentously active ingredients in a good solvent and a co-solvent; Step (4) of uniformly mixing the medicament active ingredient solution obtained in step (3) with the molten cyclopeptide of step (1); (5) subjecting the mixture obtained in step (4) to the temperature set in step (2) and subjecting the solvent to rotary evaporation under reduced pressure; The mixture obtained in step (5) is transferred to an annealing furnace and annealed at a temperature above the glass transition temperature (T ｇ ) ±50 to 150 K, preferably T ｇ (6) lowering the temperature by 50 to 100K and setting the annealing time to 30 min to 2 h, preferably 30 min to 1 h; Optionally, a step (7) of preparing a pharmaceutical composition using both the glass obtained in step (2) and a pharmaceutical adjuvant.

9. A method for producing the pharmaceutical composition of claim 4, comprising the steps of: Step (1) of dissolving the cyclopeptide and other medicamentously active ingredients in a good solvent and a co-solvent; The mixed solution obtained in step (1) is heated in an inert gas atmosphere, the solvent is rotary evaporated under reduced pressure, and then the heated temperature is kept at a temperature equal to the melting point temperature (T ｍ ) ±50 to 250 K, preferably T ｍ (2) setting the temperature at 50 to 100 K higher than the temperature at which the temperature is maintained, and setting the temperature maintenance time at 0 min to 30 h, preferably 15 to 30 min; The mixture obtained in step (5) is transferred to an annealing furnace and annealed at a temperature above the glass transition temperature (T ｇ ) ±50 to 150 K, preferably T ｇ (6) lowering the temperature by 50 to 100K and setting the annealing time to 30 min to 2 h, preferably 30 min to 1 h; Optionally, a step (7) of preparing a pharmaceutical composition using both the glass obtained in step (2) and a pharmaceutical adjuvant.

10. The pharmaceutical composition according to claim 4, wherein the glass form of the composition is further manufactured into an oral agent, a patch, a subcutaneous implant, a stent material and a microneedle device.

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