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 pharmaceutical technical field and relates to a novel dosage form and a novel adjuvant for cyclopeptide drugs, particularly to cyclopeptide glass and cyclopeptide pharmaceutical composition glass. It also relates to a manufacturing method for converting poorly soluble cyclopeptide drugs into a glass-like form by effectively utilizing related technologies for poorly soluble drugs. This cyclopeptide glass is useful in the drug delivery fields of antitumor, antiviral / antibacterial, blood sugar control, immunomodulation, neuromodulation, etc., especially in sustained drug release therapy. [Background technology]

[0002] Cyclopeptides are composed of multiple amino acids fused together via peptide bonds, and their structure has certain conformational constraints. Due to their unique topology, cyclopeptides are highly stable against chemical, thermal, and biodegradation. Cyclopeptides contain a large number of hydrogen bond acceptors and donors, and hydrogen bonding is one of the main modes of drug-receptor interaction. Therefore, cyclopeptides often possess certain biological and pharmacological activities, making them important pharmacophores in medicinal chemistry. Furthermore, due to their restricted conformation, cyclopeptides exhibit superior specificity and target affinity compared to linear peptides, resulting in a wide range of biological and pharmacological activities, including antitumor, antiviral, antibacterial, antiaging, immunomodulatory, memory enhancement, and blood glucose regulation.

[0003] However, due to their highly rigid structure and extremely strong hydrogen-bonding ability, cyclopeptide molecules suffer from poor water solubility, tendency to crystallize, and low bioavailability, which severely impact their medicinal activity. To address these shortcomings, researchers at home and abroad have adopted a variety of pharmaceutical strategies, including direct chemical modification of cyclopeptides (e.g., N-methylation) or encapsulation in nanocarriers (e.g., lipids and micelles). However, complex synthetic processes, potential toxicity, and low loading capacity of cyclopeptides remain.

[0004] Glassy structures are metastable amorphous structures that retain the biological and pharmacological activity of cyclopeptide molecules compared to more thermodynamically stable crystals. Glassy cyclopeptide drugs exhibit higher surface free energy and higher dispersity than 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, and 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 those based on pharmaceutical compositions with biological or pharmacological activity or sustained-release functions, and no methods for converting poorly soluble cyclopeptides into glassy forms.

[0007] Surprisingly, the present invention has found that cyclopeptides and their derivatives can be processed into cyclopeptide glass and cyclopeptide pharmaceutical composition glass through 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 primary objective of the present invention is to provide a cyclopeptide glass, a cyclopeptide pharmaceutical composition glass, and a method for converting a poorly soluble cyclopeptide to a glassy state. In the present invention, the term "glass" refers to an amorphous solid that exhibits glass transition, and the term "glassy" refers to a disordered structure that retains glass-like properties, lacks crystalline structure as detected by X-ray diffraction, 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 glassy state, which effectively improves the in vitro dissolution rate and solubility of poorly soluble cyclopeptides and improves the bioavailability of poorly soluble cyclopeptide drugs. [Means for solving the problem]

[0010] In a first aspect, the glass is based on a cyclopeptide, wherein the cyclopeptide is a cyclopeptide represented by structural formula 1 and / or a salt thereof, and the cyclopeptide is one type or a combination of two or more types. [ka] (A1 to A n are independently one or a combination of more than one 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, selenium cysteine, 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, and A1 to A n are condensed and linked by amino acids.)

[0011] Preferably, the cyclopeptide has biological activity and / or pharmacological activity, more preferably, antibacterial / antiviral, antitumor, blood glucose regulating, or immunomodulating activity, and antibacterial / 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 solely from cyclopeptides and their salts, or that the glasses also contain other pharmaceutical adjuvants and / or pharmaceutical active ingredients.

[0024] This pharmaceutical composition glass comprises one or more components selected from the group consisting of a cyclopeptide or a cyclopeptide derivative, another pharmaceutically active ingredient or a pharmaceutically acceptable salt thereof, and a pharmaceutical adjuvant, and preferably contains at least one active cyclopeptide; The active pharmaceutical ingredient comprises one or a mixture of two or more non-cyclopeptide drug molecules having antibacterial / antiviral, antitumor, blood glucose regulating, immunomodulating, antipsychotic, etc. effects. 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 glycoside, 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 the other pharmaceutically active ingredient is 1 to 20%, preferably 1 to 10%; The mass ratio of the pharmaceutical adjuvant is 0 to 50%, preferably 0 to 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 materials 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, and then transferring it to an annealing furnace for annealing treatment, thereby obtaining a cyclopeptide glass drug or a cyclopeptide glass adjuvant; The temperature rise 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 treatment 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, there is provided a method for producing a poorly soluble cyclopeptide pharmaceutical composition glass, the method comprising: (1) mixing the poorly soluble cyclopeptide and other pharmaceutically active ingredients, and then thoroughly ball milling the mixture while adjusting the temperature of the raw materials during ball milling to 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 using both the glass obtained in step (2) and a pharmaceutical adjuvant by one or a combination of two or more of tabletting, wet granulation, fluidized bed granulation, coating, spray granulation, program casting, and 3D printing.

[0028] Alternatively, the following manufacturing method is provided, which mainly comprises: (1) mixing the poorly soluble cyclopeptide and other pharmaceutically active ingredients, and then thoroughly ball 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 in step (1) is heated in an inert gas atmosphere and kept at the temperature for a predetermined time. The temperature is then increased to the melting point temperature (T m ) ±50 to 250 K, preferably T m Step (2) of heating the temperature to 50 to 100 K higher than the reference temperature and keeping the temperature for 0 min to 30 h, preferably 15 to 30 min; Step (3) of dissolving other pharmaceutically active ingredients in a good solvent and a co-solvent, wherein the good solvent is preferably ethanol, hexafluoroisopropanol, methylene chloride, carbon tetrachloride, acetone, or ethyl acetate, and the co-solvent is preferably one or a mixture of two or more of sodium benzoate, dimethylacetamide, urea, and Tween; Step (4) of uniformly mixing the pharmaceutically 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 Step (6) of lowering the temperature by 50 to 100K and setting the annealing treatment 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 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) is a step of dissolving the poorly soluble cyclopeptide and other pharmaceutically 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 then the heated temperature is kept at the melting point temperature (T m ) ±50 to 250 K, preferably T m Step (2) of heating the temperature to 50 to 100 K higher than the reference temperature and keeping the temperature for 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 Step (3) of setting the temperature at 50 to 100K lower than the reference temperature 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 using both the glass obtained in step (3) and the pharmaceutical adjuvant by one or a combination of two or more of tabletting, wet granulation, fluidized bed granulation, coating, spray granulation, program casting, and 3D printing.

[0030] In a sixth aspect, according to the cyclopeptide glass drug, cyclopeptide glass adjuvant, cyclopeptide pharmaceutical composition glass, and 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, cyclopeptide pharmaceutical composition glass, and 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 drug bioavailability, and reduce drug side effects. (3) It has good biocompatibility and biodegradability. (4) High thermal and chemical stability.

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

[0034] The raw materials for the cyclopeptide glass of the present invention include any one or more combinations of cyclopeptides, particularly biologically and pharmacologically active cyclopeptides or their pharmaceutically 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 a glass form, including a method for producing a glass from a poorly soluble cyclopeptide and a method for producing a glass using both a poorly soluble cyclopeptide and another active drug component. The present invention also provides a method for converting poorly soluble drugs into a glass form, forming a drug glass dosage form, and achieving poorly soluble active cyclopeptides and sustained drug release. The resulting cyclopeptide glass is highly biocompatible, biodegradable, and thermally and chemically stable. Furthermore, it can be fabricated into oral dosage forms, patches, subcutaneous implants, stent materials, and microneedle devices using techniques such as spray drying, solid dispersion, and additive manufacturing. The resulting cyclopeptide glass can exert the effects of drugs and function as a drug adjuvant. Compared with crystals and conventional pharmaceutical dosage forms or adjuvants, cyclopeptide glass can effectively accelerate the drug dissolution rate and improve drug bioavailability, making it widely applicable in the fields of drug delivery and sustained release, including antitumor, antiviral / antibacterial, blood sugar control, immunomodulation, and neuromodulation. [Brief explanation of the drawings]

[0035] [Figure 1] 1A shows an actual image of the CsA glass produced in Example 1 at room temperature, and FIG. 1B shows an actual image of the CsA glass obtained by the casting process, demonstrating the processability of cyclopeptide glass. [Figure 2] This is the nuclear magnetic resonance hydrogen spectrum of the CsA glass produced in Example 1. No significant changes were observed in the peaks compared to the CsA raw material, demonstrating that the chemical composition of the cyclopeptide raw material molecules remains unchanged even after heat melting and annealing treatments. [Figure 3]1 is a differential scanning calorimetry (DSC) diagram of the CsA glass prepared in Example 1, demonstrating the glassy structure of the CsA glass, and the glass transition temperature of the CsA glass is Tg=100.4K. [Figure 4] 1 shows the X-ray diffraction (XRD) pattern of the CsA glass produced in Example 1, demonstrating that the CsA glass has an amorphous structure. [Figure 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 sustained drug release is possible. [Figure 7] 1 shows immune-related indicators of mice orally administered with the CsA glass produced in Example 1. [Figure 8] 1 shows the XRD spectrum of the pharmaceutical composition glass based on CFP and the tumor chemotherapy drug PTX in Example 2, which has an amorphous structure. [Figure 9] The glass capsules of the pharmaceutical composition prepared in Example 2 were degraded in artificial gastric juice (prepared according to the method described in the Chinese Pharmacopoeia), demonstrating the degradability of the glass capsules of the pharmaceutical composition. [Figure 10] 1 shows the in vivo drug-time curve of mice intragastrically administered with the pharmaceutical composition glass capsules prepared in Example 2. [Figure 11] 1 shows the tumor inhibition curve of experimental mice orally administered with the pharmaceutical composition glass capsules prepared in Example 2, demonstrating the antitumor effect of the pharmaceutical composition glass capsules. [Figure 12] 1 shows the weight change curve of experimental mice orally administered with the pharmaceutical composition glass capsules prepared in Example 2, demonstrating high biosecurity. [Figure 13] 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 that the crystals were converted to a glassy state. [Figure 15] 1 shows the results of a biocompatibility test of the pharmaceutical composition glass prepared in Example 3, demonstrating high biocompatibility. [Figure 16] 1 shows the bacterial growth inhibition curve of the pharmaceutical composition glass coating prepared in Example 3. [Figure 17] 1 shows the inhibitory effect of the glass tablets of the pharmaceutical composition 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 tablets of the pharmaceutical composition prepared in Example 4, demonstrating that the glass tablets can effectively control the blood glucose of mice. [Figure 20] This is a time-dependent change curve of mouse weight when the pharmaceutical composition glass tablet prepared in Example 4 was administered intragastrically to mice with idiopathic type II diabetes, demonstrating that the mouse weight can be effectively controlled. [Figure 21] Thirty days after intragastric administration of the glass tablets 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 release rate of CLP-active cyclopeptides. [Figure 24] 1 shows the XRD pattern of the pharmaceutical composition glass prepared in Example 5, where the pharmaceutical composition glass is amorphous. [Figure 25] These are statistical data on spontaneous activity and anxiety-like behavior in mice in an open field experiment. Compared to normal mice, early social isolation mice showed a significant decrease in activity distance in the central area, but treatment with glass microneedle patches showed a significant improvement. [Figure 26] In a maze experiment, the number of entries into the open and closed arms and the time spent in the two arms were measured within 5 minutes. Compared to normal mice, the early social isolation mice showed a significant decrease in the number of entries into the open arms and a shorter time spent in the open arms, but significant improvements were observed after treatment with the glass microneedle patch. DETAILED DESCRIPTION OF THE INVENTION

[0036] The technical solutions of the present invention will be described in detail below through examples, but the scope of the protection of the present 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) CsA powder was placed in a ball mill and crushed uniformly. The ball mill was maintained at 25°C, and then the powder was transferred to a crucible. (2) The crucible containing the CsA powder in step (1) was placed in a heating device under a N2 atmosphere. (3) Heat the device of step (2), i.e., 10Kmin -1 The temperature of the crucible was raised from room temperature to 573.15 K at a rate of 1000 K, and the temperature was maintained at this temperature for 20 minutes. (4) The temperature of the device in step (3) is lowered, i.e., 10 Kmin -1 The temperature of the crucible was lowered to 273.15 K at a rate of 1000 kJ / min, 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 the casting process, demonstrating the processability of cyclopeptide glass.

[0039] FIG. 2 shows the nuclear magnetic resonance hydrogen spectrum of the CsA glass produced in Example 1. No significant changes were observed in the peaks compared to the CsA raw material, demonstrating that the chemical composition of the cyclopeptide raw material molecules remains unchanged even after heat melting and annealing treatments.

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

[0041] FIG. 4 shows the 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, demonstrating that sustained drug release is possible. 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 the immune-related indexes of mice orally administered the CsA glass prepared in Example 1. Specifically, healthy male Kunming mice (weight 20±1 g, 4-5 weeks old) were randomly divided into two groups of eight mice each and fed a normal diet. The mice in the experimental group received 40 mg kg -1The mice were intragastrically administered a total of five doses of 100 mg ... + (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 maintained at 37°C, and then the mixture was transferred to a container. (2) Heat the vessel of step (1), i.e., 20 Kmin -1 The temperature of the container was raised from room temperature to 473.15 K at a rate of 100°C, and the container was kept at this temperature for 30 minutes. (3) The temperature of the vessel in step (2) is lowered, i.e., 10 K min -1 The temperature of the vessel was lowered to 333.15 K at a rate of 0.05°C. (4) The PTX powder was placed in a ball mill and crushed uniformly. The ball mill was maintained 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., 10 Kmin -1The container was cooled to 315.15 K at a cooling rate of 0.055 and kept at that temperature for 30 min in an annealing furnace to obtain a pharmaceutical composition glass based on CFP and the tumor chemotherapy drug PTX. The glass was completely dissolved in an HFIP organic solution, and the PTX concentration was quantified by high-performance liquid chromatography. The PTX loading was calculated to be 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 an 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 capsules prepared in Example 2 in artificial gastric fluid (according to the manufacturing method prescribed in the Chinese Pharmacopoeia), demonstrating the degradability of the pharmaceutical composition glass capsules. The manufacturing steps for the pharmaceutical composition glass capsules are to crush the pharmaceutical composition glass into powder using a homogenizer, and then fill the powder into gelatin capsule shells.

[0048] Figure 10 shows the in vivo drug-time curve of mice intragastrically administered with the pharmaceutical composition glass capsules prepared in Example 2. Specifically, mice were administered 8 mg kg -1 After intragastric administration at a dose of , 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] Figure 11 shows the tumor inhibition curve of experimental mice orally administered with the pharmaceutical composition glass capsules prepared in Example 2, demonstrating the antitumor effect of the pharmaceutical composition glass capsules. 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 breast cancer mouse transplant tumor model. The tumors in the mice were 80±10 mm 3After the mice had grown to a volume of 8 mg kg, they were treated with the drug. -1 The mice in the control group were given the same amount of PBS every other day. The changes in 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 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 changes in organ indexes 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 day 35 and collected. The surface blood was flushed out with PBS, and the organs were absorbed with filter paper. They were then weighed on an electronic balance, and the organ weight (mg) was divided by the total weight (g) of the mice to calculate the organ index.

[0050] Example 3 The method for producing a 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, with the mass ratio of these three being 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) The mixed solution obtained in step (2) was placed in a vacuum rotary evaporator and evaporated for 50 Kmin. -1 The temperature was raised from room temperature to 443.15 K at a 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., 10 Kmin -1 The temperature was lowered to 243.15 K at a rate of 0.05°C and kept at this temperature for 20 minutes. (5) To the product obtained in step (4), a pharmaceutical adjuvant containing 1.5 parts starch, 1 part sodium carboxymethylcellulose, 2.5 parts polyethylene glycol and 6.5 parts hydrogenated vegetable oil was added in a total amount of 92.5% by weight. (6) The pharmaceutical composition obtained in step (5) was homogenized, and glass pharmaceutical composition embeddings were prepared by a combined technique of tableting and vacuum spray drying.

[0051] FIG. 14 shows the 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 for the pharmaceutical composition glass prepared in Example 3, demonstrating high biocompatibility. Specifically, the glass was processed using a coater to form a cell culture dish coating with a diameter of 60 mm and a thickness of 0.2 mm, and used for cell culture (fibroblast 3T3). After 48 hours of incubation, 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 1 cm x 1 cm x 0.1 cm tablet and attached to the center of a bacterial culture dish. The total drug concentration was calculated to be 10 μg / dish. E. coli and Staphylococcus aureus were uniformly mixed in a liquid medium, then added dropwise to the bacterial culture dish and incubated with shaking at 37°C. The bacterial concentration in the bacterial culture dish was measured over time using the OD value method.

[0054] Figure 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, and an Oxford cup (diameter 6 mm) was placed on an agar culture dish. The tablet was then placed in the Oxford cup and allowed to adhere to the agar. E. coli and Staphylococcus aureus were uniformly mixed with PBS, then applied to 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 (weight 20±1 g, 4-5 weeks old) were randomly divided into two groups of 15 mice each. A 1 cm window was surgically opened in the back of the mice in the experimental group, and 30±2.5 g of the pharmaceutical composition glass was subcutaneously implanted and sutured. The mice were sampled over time, the pharmaceutical composition glass was removed, and the remaining mass was weighed, and a decomposition curve was plotted.

[0056] Figure 19 shows the blood glucose control curve of mice with idiopathic type II diabetes using the glass tablets of the pharmaceutical composition 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, while the control group received the same amount of saline intragastrically. The test period was 30 days. Tail blood was collected every day, and the fasting blood glucose levels of the mice 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] Figure 20 shows the time course curve of the body weight of mice when the pharmaceutical composition glass tablets prepared in Example 4 were intragastrically administered to mice with idiopathic type II diabetes, 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 concentrations of glucose are orally administered, and a curve was plotted by measuring the blood glucose concentration in the sole blood of the mice, demonstrating that the experimental group mice had higher glucose tolerance.

[0059] Figure 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 release 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 added to the above-mentioned release solution, stirred, and incubated, and the concentration of CLP in the release solution was detected.

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

[0062] Figure 25 shows statistical data on the 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. Compared to normal mice, early social isolation mice showed a significantly reduced central area activity distance, but treatment with glass microneedle patches showed a significant improvement. Specifically, C57BL / 6 early social isolation mice (8 weeks old, male) were divided into two groups of an average of six mice each. One group received no special treatment, while the other group received hair removal on their backs followed by application of glass microneedle patches. The C57BL / 6 group consisted of six normal mice and served as a blank control. According to the standard, a black open field apparatus (45 cm × 5 cm × 45 cm) was prepared, and mice were placed in the open field. The spontaneous activity of the mice was tracked and recorded. The ordinate represents the statistical value of central area activity distance during 1 h of open field activity.

[0063] Figure 26 shows the number of entries into the open and closed arms and the time spent in the two arms of mice within 5 minutes when the pharmaceutical glass composition prepared in Example 6 was used in a mouse maze experiment. Compared to normal mice, early social isolation mice showed a significantly reduced number of entries into the open arms and a shorter time spent in the open arms, but treatment with the glass microneedle patch showed significant improvement. Specifically, a maze apparatus was prepared according to the standard: arm width 5 cm, arm length 35 cm, closed arm height 15 cm, central platform 5 cm x 5 cm, and maze height from the ground approximately 40-55 cm. Mice were placed in the center of the platform, and the number of entries into the open and closed arms and the time spent in the two arms were recorded to evaluate anxiety indices.

[0064] Example 4 The method for preparing a pharmaceutical composition glass based on histidine-proline cyclodipeptide and secretide comprises the following steps: (1) Histidine-proline cyclodipeptide and secretide were placed in a ball mill, pulverized, and mixed uniformly. The ball mill was maintained at 25°C, and then transferred to a container, and the mass ratio of the two was adjusted to 10:1. (2) Dichloromethane solvent was added to the mixed powder of step (1) and stirred until completely dissolved. (3) The mixed solution obtained in step (2) was placed in a vacuum rotary evaporator and evaporated for 10 Kmin. -1 The temperature was raised from room temperature to 535.15 K at a 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., 10 Kmin -1 The temperature was lowered to 283.15 K at a rate of 100°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 equimolar ratios and dissolved in hexafluoroisopropanol to form a single cyclopeptide solution. (2) The mixed solution obtained in step (1) was placed in a vacuum rotary evaporator and evaporated for 10 Kmin. -1 The temperature was raised from room temperature to 473.15 K at a rate of 100°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., 10 Kmin -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 comprises the following steps: (1) CWW and risperidone powder were weighed in an equimolar ratio and dissolved in hexafluoroisopropanol and methanol, respectively, to form monodisperse solutions. (2) The mixed solution obtained in step (1) was placed in a vacuum rotary evaporator and evaporated for 10 Kmin. -1 The temperature was raised from room temperature to 573.15 K at a rate of 100°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., 50 Kmin -1 The temperature was lowered to 283.15 K at a rate of 0.15°C and 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 measuring 0.8 × 0.8 cm by 3D printing and casting processes and used for transdermal drug 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 100 mg of schizophrenia-like steroids and evaluated for schizophrenia-like behavior after 14 days. The inventions described in the claims of the original application are as follows: [1] 1. A cyclopeptide-based glass comprising: The cyclopeptide is a cyclopeptide represented by structural formula 1 and a salt thereof, and may be one type or a combination of two or more types. The cyclopeptide-based glass is characterized in that the cyclopeptide preferably has biological activity and / or pharmacological activity, and more preferably has antibacterial, antiviral, antitumor, blood glucose regulating, and immunomodulating activity. [ka] ( 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, selenium cysteine, 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, and t-butoxycarbonyl. , n≧2, preferably 2≦n≦15, and A 1 ~A n are condensed and linked by amino acids.) [2] The antibacterial and antiviral cyclopeptide is one or a combination of two or more selected from the following cyclopeptides: [Table 7-1] [Table 7-2] The antitumor cyclopeptide is one or a combination of multiple cyclopeptides selected from the following: [Table 8-1] [Table 8-2] The immunomodulatory cyclopeptide is one or a combination of multiple cyclopeptides selected from the following: [Table 9] The blood glucose-regulating cyclopeptide is one or a combination of multiple cyclopeptides selected from the following:

Table 10

Table 11

Table 12

[10] The pharmaceutical composition according to [4], wherein the glass form of the composition is further manufactured into oral preparations, patches, subcutaneous implants, stent materials, and microneedle devices.

Claims

1. A cyclopeptide glass comprising: The cyclopeptide is one or a combination of two or more cyclopeptides, The cyclopeptide has antibacterial / antiviral activity, antitumor activity, immunomodulatory activity, blood glucose regulating activity, cardiovascular and hematological activity, or other activity; The cyclopeptide having antibacterial and antiviral activity is one or a combination of two or more selected from the following cyclopeptides: 【Table 1-1】 【Table 1-2】 The cyclopeptide having antitumor activity is one or a combination of two or more selected from the following cyclopeptides: 【Table 2-1】 【Table 2-2】 The cyclopeptide having immunomodulatory activity is one or a combination of two or more selected from the following cyclopeptides: 【Table 3】 The cyclopeptide having blood glucose-regulating activity is one or a combination of two or more selected from the following cyclopeptides: 【Table 4】 The cyclopeptide having cardiovascular and blood-related activity is one or a combination of several cyclopeptides selected from the following: 【Table 5】 The cyclopeptides having other activities are one or a combination of two or more selected from the following cyclopeptides: 【Table 6】 Cyclopeptide glass.

2. The cyclopeptide glass according to claim 1, wherein 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.

3. A pharmaceutical composition containing the cyclopeptide glass according to claim 1 or 2, The pharmaceutical composition is in glass form and is made solely of cyclopeptides or further contains pharmaceutical adjuvants and / or pharmaceutical active ingredients.

4. A method for producing the cyclopeptide glass according to claim 1 or 2, comprising: (1) controlling the temperature of the raw material during ball milling to 0 to 50°C, and ball milling one or more cyclopeptides; and (2) producing cyclopeptide glass by a "heating-quenching" method, in which the crushed cyclopeptide raw material is heated to a melting point temperature (T m ) ±50 to 250 K in an inert gas atmosphere, kept at that temperature for a predetermined period of time, and then transferred to an annealing furnace for annealing.

5. The temperature rise is the melting point temperature (T ｍ ) ±50 to 250K The warming time is 0 min to 30 h. The annealing temperature is the glass transition temperature (T ｇ ) ±50 to 150 Kelvin The annealing time is 30 min to 2 h. T ｍ and T ｇ is measured by thermogravimetric analysis and differential scanning calorimetry, and the temperature rise and fall rates are 2 to 50 km / min. －１ 5. The method of claim 4, wherein:

6. A method for producing the pharmaceutical composition of claim 3, comprising: (1) mixing the cyclopeptide with other pharmaceutically active ingredients and ball milling the mixture while controlling the temperature of the raw materials during ball milling to 0-50°C; Step (2) of producing cyclopeptide glass by the "heating-quenching" method, in which the crushed cyclopeptide raw material is heated to a melting point temperature (T m ) ±50 to 250 K in an inert gas atmosphere, kept at that temperature for a predetermined time, and then transferred to an annealing furnace for annealing to obtain glass; and optionally, step (3) of preparing a pharmaceutical composition using both the glass obtained in step (2) and a pharmaceutical adjuvant.

7. A method for producing the pharmaceutical composition of claim 3, comprising: (1) controlling the temperature of the raw material during ball milling to 0 to 50°C, and ball milling one or more cyclopeptides; The powder in step (1) is heated in an inert gas atmosphere and kept at the temperature for a predetermined time. The temperature is then increased to the melting point temperature (T ｍ ) ±50 to 250 K, and the temperature is kept at 0 min to 30 h. Step (3) of dissolving other pharmaceutically active ingredients in a good solvent and a co-solvent; Step (4) of uniformly mixing the pharmaceutically active ingredient solution obtained in step (3) with the molten cyclopeptide of step (1); Step (5) of subjecting the mixture obtained in step (4) to the temperature set in step (2) and rotary evaporating the solvent 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, and annealing for 30 min to 2 h (step (6)); Optionally, a step (7) 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 3, comprising: Step (1) of dissolving the cyclopeptide and other pharmaceutically 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 evaporated by rotary evaporation under reduced pressure, and then the heated temperature is kept at a temperature equivalent to the melting point temperature (T ｍ ) ±50 to 250 K, and the temperature is kept at 0 min to 30 h. 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, and annealing for 30 min to 2 h; Optionally, a step (4) of preparing a pharmaceutical composition using both the glass obtained in step (3) and a pharmaceutical adjuvant.

9. The pharmaceutical composition of claim 3, wherein the glass form of the composition is further manufactured into oral agents, patches, subcutaneous implants, stent materials, and microneedle devices.

Images