CN117045624A - Wogonin inhalation preparation and preparation method and medical application thereof - Google Patents
Wogonin inhalation preparation and preparation method and medical application thereof Download PDFInfo
- Publication number
- CN117045624A CN117045624A CN202311064407.6A CN202311064407A CN117045624A CN 117045624 A CN117045624 A CN 117045624A CN 202311064407 A CN202311064407 A CN 202311064407A CN 117045624 A CN117045624 A CN 117045624A
- Authority
- CN
- China
- Prior art keywords
- wogonin
- inhalation
- preparation
- solution
- lung
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
- HIMJIPRMECETLJ-UHFFFAOYSA-N Wogonin Natural products COc1cc(O)c(O)c2C(=O)C=C(Oc12)c3ccccc3 HIMJIPRMECETLJ-UHFFFAOYSA-N 0.000 title claims abstract description 98
- XLTFNNCXVBYBSX-UHFFFAOYSA-N wogonin Chemical compound COC1=C(O)C=C(O)C(C(C=2)=O)=C1OC=2C1=CC=CC=C1 XLTFNNCXVBYBSX-UHFFFAOYSA-N 0.000 title claims abstract description 97
- 238000002360 preparation method Methods 0.000 title claims abstract description 45
- 229940041682 inhalant solution Drugs 0.000 claims abstract description 31
- 208000005069 pulmonary fibrosis Diseases 0.000 claims abstract description 19
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 claims abstract description 18
- 206010058467 Lung neoplasm malignant Diseases 0.000 claims abstract description 11
- 201000005202 lung cancer Diseases 0.000 claims abstract description 11
- 208000020816 lung neoplasm Diseases 0.000 claims abstract description 11
- 206010069351 acute lung injury Diseases 0.000 claims abstract description 9
- 239000011780 sodium chloride Substances 0.000 claims abstract description 8
- 239000007788 liquid Substances 0.000 claims abstract description 4
- 239000003814 drug Substances 0.000 claims description 24
- 239000004475 Arginine Substances 0.000 claims description 17
- ODKSFYDXXFIFQN-UHFFFAOYSA-N arginine Natural products OC(=O)C(N)CCCNC(N)=N ODKSFYDXXFIFQN-UHFFFAOYSA-N 0.000 claims description 17
- 230000000694 effects Effects 0.000 claims description 13
- 239000000203 mixture Substances 0.000 claims description 11
- 238000000034 method Methods 0.000 claims description 10
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 10
- 238000009472 formulation Methods 0.000 claims description 9
- ODKSFYDXXFIFQN-BYPYZUCNSA-P L-argininium(2+) Chemical compound NC(=[NH2+])NCCC[C@H]([NH3+])C(O)=O ODKSFYDXXFIFQN-BYPYZUCNSA-P 0.000 claims description 3
- 238000007906 compression Methods 0.000 claims description 3
- 230000006835 compression Effects 0.000 claims description 3
- XUJNEKJLAYXESH-REOHCLBHSA-N L-Cysteine Chemical compound SC[C@H](N)C(O)=O XUJNEKJLAYXESH-REOHCLBHSA-N 0.000 claims description 2
- HNDVDQJCIGZPNO-YFKPBYRVSA-N L-histidine Chemical compound OC(=O)[C@@H](N)CC1=CN=CN1 HNDVDQJCIGZPNO-YFKPBYRVSA-N 0.000 claims description 2
- KDXKERNSBIXSRK-YFKPBYRVSA-N L-lysine Chemical compound NCCCC[C@H](N)C(O)=O KDXKERNSBIXSRK-YFKPBYRVSA-N 0.000 claims description 2
- KDXKERNSBIXSRK-UHFFFAOYSA-N Lysine Natural products NCCCCC(N)C(O)=O KDXKERNSBIXSRK-UHFFFAOYSA-N 0.000 claims description 2
- 239000004472 Lysine Substances 0.000 claims description 2
- 238000009835 boiling Methods 0.000 claims description 2
- XUJNEKJLAYXESH-UHFFFAOYSA-N cysteine Natural products SCC(N)C(O)=O XUJNEKJLAYXESH-UHFFFAOYSA-N 0.000 claims description 2
- 235000018417 cysteine Nutrition 0.000 claims description 2
- 239000012467 final product Substances 0.000 claims description 2
- 238000010438 heat treatment Methods 0.000 claims description 2
- HNDVDQJCIGZPNO-UHFFFAOYSA-N histidine Natural products OC(=O)C(N)CC1=CN=CN1 HNDVDQJCIGZPNO-UHFFFAOYSA-N 0.000 claims description 2
- 238000002604 ultrasonography Methods 0.000 claims description 2
- 239000003595 mist Substances 0.000 claims 1
- 210000004072 lung Anatomy 0.000 abstract description 26
- 239000002245 particle Substances 0.000 abstract description 5
- 239000000126 substance Substances 0.000 abstract description 3
- 238000013461 design Methods 0.000 abstract description 2
- 150000001413 amino acids Chemical class 0.000 abstract 1
- 239000006184 cosolvent Substances 0.000 abstract 1
- 241000699670 Mus sp. Species 0.000 description 28
- 239000000443 aerosol Substances 0.000 description 20
- 239000000243 solution Substances 0.000 description 18
- 238000002474 experimental method Methods 0.000 description 15
- 241000699666 Mus <mouse, genus> Species 0.000 description 11
- 229940079593 drug Drugs 0.000 description 10
- KRKNYBCHXYNGOX-UHFFFAOYSA-N citric acid Chemical compound OC(=O)CC(O)(C(O)=O)CC(O)=O KRKNYBCHXYNGOX-UHFFFAOYSA-N 0.000 description 9
- VEXZGXHMUGYJMC-UHFFFAOYSA-N Hydrochloric acid Chemical compound Cl VEXZGXHMUGYJMC-UHFFFAOYSA-N 0.000 description 8
- 239000012530 fluid Substances 0.000 description 8
- 229930040373 Paraformaldehyde Natural products 0.000 description 7
- 229920002866 paraformaldehyde Polymers 0.000 description 7
- 108010006654 Bleomycin Proteins 0.000 description 6
- 238000010171 animal model Methods 0.000 description 6
- 229960001561 bleomycin Drugs 0.000 description 6
- OYVAGSVQBOHSSS-UAPAGMARSA-O bleomycin A2 Chemical compound N([C@H](C(=O)N[C@H](C)[C@@H](O)[C@H](C)C(=O)N[C@@H]([C@H](O)C)C(=O)NCCC=1SC=C(N=1)C=1SC=C(N=1)C(=O)NCCC[S+](C)C)[C@@H](O[C@H]1[C@H]([C@@H](O)[C@H](O)[C@H](CO)O1)O[C@@H]1[C@H]([C@@H](OC(N)=O)[C@H](O)[C@@H](CO)O1)O)C=1N=CNC=1)C(=O)C1=NC([C@H](CC(N)=O)NC[C@H](N)C(N)=O)=NC(N)=C1C OYVAGSVQBOHSSS-UAPAGMARSA-O 0.000 description 6
- 239000002158 endotoxin Substances 0.000 description 6
- 229920006008 lipopolysaccharide Polymers 0.000 description 6
- 102000004169 proteins and genes Human genes 0.000 description 6
- 108090000623 proteins and genes Proteins 0.000 description 6
- 206010016654 Fibrosis Diseases 0.000 description 5
- 230000004761 fibrosis Effects 0.000 description 5
- 238000010172 mouse model Methods 0.000 description 5
- 235000018102 proteins Nutrition 0.000 description 5
- 210000002966 serum Anatomy 0.000 description 5
- 241001465754 Metazoa Species 0.000 description 4
- 238000000889 atomisation Methods 0.000 description 4
- 238000010586 diagram Methods 0.000 description 4
- 201000010099 disease Diseases 0.000 description 4
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 4
- 230000002757 inflammatory effect Effects 0.000 description 4
- 239000000463 material Substances 0.000 description 4
- 239000012188 paraffin wax Substances 0.000 description 4
- 239000002994 raw material Substances 0.000 description 4
- 239000008215 water for injection Substances 0.000 description 4
- -1 5, 7-dihydroxy-8-methoxy-2-phenyl-4H-1-benzopyran-4-one (5, 7-dihydroxy-8-methoxy flavone) Chemical compound 0.000 description 3
- 102000004127 Cytokines Human genes 0.000 description 3
- 108090000695 Cytokines Proteins 0.000 description 3
- 238000002965 ELISA Methods 0.000 description 3
- 102000004889 Interleukin-6 Human genes 0.000 description 3
- 108090001005 Interleukin-6 Proteins 0.000 description 3
- 208000004852 Lung Injury Diseases 0.000 description 3
- 206010069363 Traumatic lung injury Diseases 0.000 description 3
- 230000037396 body weight Effects 0.000 description 3
- 210000000621 bronchi Anatomy 0.000 description 3
- 239000003153 chemical reaction reagent Substances 0.000 description 3
- 230000008021 deposition Effects 0.000 description 3
- 238000004128 high performance liquid chromatography Methods 0.000 description 3
- 239000007924 injection Substances 0.000 description 3
- 238000002347 injection Methods 0.000 description 3
- 210000000265 leukocyte Anatomy 0.000 description 3
- 231100000515 lung injury Toxicity 0.000 description 3
- 230000003285 pharmacodynamic effect Effects 0.000 description 3
- 238000011160 research Methods 0.000 description 3
- 238000007619 statistical method Methods 0.000 description 3
- 238000012360 testing method Methods 0.000 description 3
- 230000001225 therapeutic effect Effects 0.000 description 3
- MZOFCQQQCNRIBI-VMXHOPILSA-N (3s)-4-[[(2s)-1-[[(2s)-1-[[(1s)-1-carboxy-2-hydroxyethyl]amino]-4-methyl-1-oxopentan-2-yl]amino]-5-(diaminomethylideneamino)-1-oxopentan-2-yl]amino]-3-[[2-[[(2s)-2,6-diaminohexanoyl]amino]acetyl]amino]-4-oxobutanoic acid Chemical compound OC[C@@H](C(O)=O)NC(=O)[C@H](CC(C)C)NC(=O)[C@H](CCCN=C(N)N)NC(=O)[C@H](CC(O)=O)NC(=O)CNC(=O)[C@@H](N)CCCCN MZOFCQQQCNRIBI-VMXHOPILSA-N 0.000 description 2
- 238000011746 C57BL/6J (JAX™ mouse strain) Methods 0.000 description 2
- 102000008186 Collagen Human genes 0.000 description 2
- 108010035532 Collagen Proteins 0.000 description 2
- 206010028980 Neoplasm Diseases 0.000 description 2
- 108060008682 Tumor Necrosis Factor Proteins 0.000 description 2
- 102000000852 Tumor Necrosis Factor-alpha Human genes 0.000 description 2
- 239000008351 acetate buffer Substances 0.000 description 2
- 238000000540 analysis of variance Methods 0.000 description 2
- 238000004458 analytical method Methods 0.000 description 2
- 230000009286 beneficial effect Effects 0.000 description 2
- 210000004369 blood Anatomy 0.000 description 2
- 239000008280 blood Substances 0.000 description 2
- 229920001436 collagen Polymers 0.000 description 2
- 238000004090 dissolution Methods 0.000 description 2
- 230000000857 drug effect Effects 0.000 description 2
- 239000000706 filtrate Substances 0.000 description 2
- 238000001914 filtration Methods 0.000 description 2
- 229930003935 flavonoid Natural products 0.000 description 2
- 235000017173 flavonoids Nutrition 0.000 description 2
- 238000007490 hematoxylin and eosin (H&E) staining Methods 0.000 description 2
- 230000005764 inhibitory process Effects 0.000 description 2
- 239000007928 intraperitoneal injection Substances 0.000 description 2
- 239000012528 membrane Substances 0.000 description 2
- 238000012544 monitoring process Methods 0.000 description 2
- 231100000915 pathological change Toxicity 0.000 description 2
- 230000036285 pathological change Effects 0.000 description 2
- 230000001575 pathological effect Effects 0.000 description 2
- 239000008363 phosphate buffer Substances 0.000 description 2
- 230000002685 pulmonary effect Effects 0.000 description 2
- 239000008213 purified water Substances 0.000 description 2
- 239000011734 sodium Substances 0.000 description 2
- 238000007910 systemic administration Methods 0.000 description 2
- 229910021642 ultra pure water Inorganic materials 0.000 description 2
- 239000012498 ultrapure water Substances 0.000 description 2
- JVDHWXLTNDKLIZ-WCCKRBBISA-N (2s)-2-amino-5-(diaminomethylideneamino)pentanoic acid;hydrate Chemical compound O.OC(=O)[C@@H](N)CCCNC(N)=N JVDHWXLTNDKLIZ-WCCKRBBISA-N 0.000 description 1
- GOZMBJCYMQQACI-UHFFFAOYSA-N 6,7-dimethyl-3-[[methyl-[2-[methyl-[[1-[3-(trifluoromethyl)phenyl]indol-3-yl]methyl]amino]ethyl]amino]methyl]chromen-4-one;dihydrochloride Chemical compound Cl.Cl.C=1OC2=CC(C)=C(C)C=C2C(=O)C=1CN(C)CCN(C)CC(C1=CC=CC=C11)=CN1C1=CC=CC(C(F)(F)F)=C1 GOZMBJCYMQQACI-UHFFFAOYSA-N 0.000 description 1
- 101150020966 Acta2 gene Proteins 0.000 description 1
- 238000011729 BALB/c nude mouse Methods 0.000 description 1
- FXNFHKRTJBSTCS-UHFFFAOYSA-N Baicalein Natural products C=1C(=O)C=2C(O)=C(O)C(O)=CC=2OC=1C1=CC=CC=C1 FXNFHKRTJBSTCS-UHFFFAOYSA-N 0.000 description 1
- 102000000589 Interleukin-1 Human genes 0.000 description 1
- 108010002352 Interleukin-1 Proteins 0.000 description 1
- 102000003777 Interleukin-1 beta Human genes 0.000 description 1
- 108090000193 Interleukin-1 beta Proteins 0.000 description 1
- 206010025066 Lung carcinoma cell type unspecified stage 0 Diseases 0.000 description 1
- 241001529936 Murinae Species 0.000 description 1
- 206010035664 Pneumonia Diseases 0.000 description 1
- 238000011529 RT qPCR Methods 0.000 description 1
- 240000004534 Scutellaria baicalensis Species 0.000 description 1
- 235000017089 Scutellaria baicalensis Nutrition 0.000 description 1
- 238000010521 absorption reaction Methods 0.000 description 1
- UDFLTIRFTXWNJO-UHFFFAOYSA-N baicalein Chemical compound O1C2=CC(=O)C(O)=C(O)C2=C(O)C=C1C1=CC=CC=C1 UDFLTIRFTXWNJO-UHFFFAOYSA-N 0.000 description 1
- 229940015301 baicalein Drugs 0.000 description 1
- UREBDLICKHMUKA-CXSFZGCWSA-N dexamethasone Chemical compound C1CC2=CC(=O)C=C[C@]2(C)[C@]2(F)[C@@H]1[C@@H]1C[C@@H](C)[C@@](C(=O)CO)(O)[C@@]1(C)C[C@@H]2O UREBDLICKHMUKA-CXSFZGCWSA-N 0.000 description 1
- 229960003957 dexamethasone Drugs 0.000 description 1
- 230000037213 diet Effects 0.000 description 1
- 235000005911 diet Nutrition 0.000 description 1
- 239000002552 dosage form Substances 0.000 description 1
- 231100000673 dose–response relationship Toxicity 0.000 description 1
- 239000003651 drinking water Substances 0.000 description 1
- 235000020188 drinking water Nutrition 0.000 description 1
- 238000012377 drug delivery Methods 0.000 description 1
- 230000036267 drug metabolism Effects 0.000 description 1
- 239000010419 fine particle Substances 0.000 description 1
- 238000002513 implantation Methods 0.000 description 1
- 238000011503 in vivo imaging Methods 0.000 description 1
- 238000011065 in-situ storage Methods 0.000 description 1
- 238000011081 inoculation Methods 0.000 description 1
- 238000011835 investigation Methods 0.000 description 1
- 238000009688 liquid atomisation Methods 0.000 description 1
- 238000005259 measurement Methods 0.000 description 1
- 230000003340 mental effect Effects 0.000 description 1
- 108020004999 messenger RNA Proteins 0.000 description 1
- 230000037230 mobility Effects 0.000 description 1
- 239000000178 monomer Substances 0.000 description 1
- 238000000465 moulding Methods 0.000 description 1
- 238000002663 nebulization Methods 0.000 description 1
- 238000001543 one-way ANOVA Methods 0.000 description 1
- 238000005457 optimization Methods 0.000 description 1
- 231100000683 possible toxicity Toxicity 0.000 description 1
- 239000000047 product Substances 0.000 description 1
- 238000003762 quantitative reverse transcription PCR Methods 0.000 description 1
- 229920006395 saturated elastomer Polymers 0.000 description 1
- 238000013424 sirius red staining Methods 0.000 description 1
- 238000010186 staining Methods 0.000 description 1
- 208000024891 symptom Diseases 0.000 description 1
- 231100000057 systemic toxicity Toxicity 0.000 description 1
- 238000010998 test method Methods 0.000 description 1
- 238000009210 therapy by ultrasound Methods 0.000 description 1
- 238000013518 transcription Methods 0.000 description 1
- 230000035897 transcription Effects 0.000 description 1
- 210000004881 tumor cell Anatomy 0.000 description 1
- 238000007492 two-way ANOVA Methods 0.000 description 1
Classifications
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K9/00—Medicinal preparations characterised by special physical form
- A61K9/0012—Galenical forms characterised by the site of application
- A61K9/007—Pulmonary tract; Aromatherapy
- A61K9/0073—Sprays or powders for inhalation; Aerolised or nebulised preparations generated by other means than thermal energy
- A61K9/0078—Sprays or powders for inhalation; Aerolised or nebulised preparations generated by other means than thermal energy for inhalation via a nebulizer such as a jet nebulizer, ultrasonic nebulizer, e.g. in the form of aqueous drug solutions or dispersions
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/33—Heterocyclic compounds
- A61K31/335—Heterocyclic compounds having oxygen as the only ring hetero atom, e.g. fungichromin
- A61K31/35—Heterocyclic compounds having oxygen as the only ring hetero atom, e.g. fungichromin having six-membered rings with one oxygen as the only ring hetero atom
- A61K31/352—Heterocyclic compounds having oxygen as the only ring hetero atom, e.g. fungichromin having six-membered rings with one oxygen as the only ring hetero atom condensed with carbocyclic rings, e.g. methantheline
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K9/00—Medicinal preparations characterised by special physical form
- A61K9/08—Solutions
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P11/00—Drugs for disorders of the respiratory system
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P35/00—Antineoplastic agents
Landscapes
- Health & Medical Sciences (AREA)
- Chemical & Material Sciences (AREA)
- General Health & Medical Sciences (AREA)
- Veterinary Medicine (AREA)
- Public Health (AREA)
- Medicinal Chemistry (AREA)
- Pharmacology & Pharmacy (AREA)
- Life Sciences & Earth Sciences (AREA)
- Animal Behavior & Ethology (AREA)
- Engineering & Computer Science (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Epidemiology (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Pulmonology (AREA)
- General Chemical & Material Sciences (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Organic Chemistry (AREA)
- Otolaryngology (AREA)
- Dispersion Chemistry (AREA)
- Medicinal Preparation (AREA)
Abstract
The invention relates to a wogonin inhalation preparation, a preparation method and medical application thereof, which comprises wogonin inhalation solution and an atomizer matched with the wogonin inhalation solution for use. The wogonin inhalation solution takes amino acid substances as cosolvent, and sodium chloride can also be added as isotonic regulator. The liquid preparation obtained by reasonable formula design has good stability and uniform particle size, and meets the requirements of lung inhalation preparations. The preparation can be used for treating acute lung injury, pulmonary fibrosis and lung cancer.
Description
Technical Field
The invention relates to a medicinal preparation, a preparation method and application thereof, in particular to a wogonin inhalation preparation, a preparation method and medical application thereof.
Background
As a commonly used traditional Chinese medicine, baikal skullcap root has been proved to be effective in treatment of various diseases from ancient times. Modern scientific research finds that wogonin (Baicalein) is an effective monomer for treating radix Scutellariae, and has chemical name of 5, 7-dihydroxy-8-methoxy-2-phenyl-4H-1-benzopyran-4-one (5, 7-dihydroxy-8-methoxy flavone) and molecular formula of C 16 H 12 O 5 (MW.284.26), the chemical structural formula is shown as formula (1). However, the study shows that wogonin itself has low bioavailability, and even in the treatment and study of lung related diseases, oral administration, injection and other modes are often adopted to make the medicine enter the body. Although the bioavailability of systemic administration can be improved by dosage form improvement, the dosage is required to be increased to ensure the drug concentration at the focus, and the systemic administration of wogonin can not only cause potential toxicity, but also greatly increase the burden of the organism of a patient on drug metabolism. The inhalation preparation is a good formulation for pulmonary administration, and can reach lung focus through simple process and convenient administration, and the capillary vessel with abundant lung has unique advantage for satisfying drug absorption of the inhalation preparation, thus being a good formulation for pulmonary related diseases. Based on the characteristics, if the wogonin is inhaled and administered in the treatment of lung related diseases, three optimizations can be achieved: (1) wogonin plays a local role in the lung, avoiding systemic toxicity; (2) the dosage of wogonin can be reduced; (3) convenient use and high acceptance, and can reduce the psychological and physiological discomfort of the patient in taking the medicine.
At present, no report exists that wogonin inhalation solution is prepared into an inhalation preparation product and is applied to the treatment of acute lung injury, pulmonary fibrosis and lung cancer.
Disclosure of Invention
The invention aims to: the invention aims to provide a wogonin inhalation preparation for improving drug effect and patient compliance. It is also an object of the present invention to provide a process for the preparation of said wogonin inhalant and its use in acute lung injury, pulmonary fibrosis and lung cancer.
The technical scheme is as follows: the wogonin inhalation preparation comprises wogonin inhalation solution and an atomizer matched with the wogonin inhalation solution for use.
The wogonin inhalation preparation comprises the following components in percentage by weight: 1-5 μm.
The wogonin inhalation preparation comprises the following components in percentage by weight: d (D) 10 0.5-1.0 μm, D 50 1.5-5.0 μm, D 90 3.0-10 μm.
The wogonin inhalation preparation comprises wogonin 0.1-5.0% (w/v), sodium chloride 0.85-0.95% (w/v) and arginine, lysine, histidine or cysteine 2-10% (w/v); the pH value is 7.0-11.0. Preferably, the pH is 9.0 to 11.0. Or citric acid may be used to adjust the pH of the inhalation solution.
The wogonin inhalation preparation, and the atomizer matched with the wogonin inhalation solution comprises a gas compression atomizer and an aerosol for realizing liquid atomization by means of a spring and a capillary tube.
The preparation method of the wogonin inhalation preparation comprises the following steps: the preparation method comprises dissolving arginine in injectable water, adding wogonin, and ultrasound, or further heating or boiling to obtain the final product.
The wogonin inhalation preparation is applied to the preparation of medicaments for treating acute lung injury, pulmonary fibrosis or lung cancer.
The application is that the daily dosage of wogonin is 50 mg-300 mg/60 kg/person of acute lung injury.
The application is that the lung fibrosis is 200 mg-800 mg/60 kg/person according to the daily dosage of wogonin.
The application is that the daily dosage of wogonin is 50 mg-300 mg/60 kg/person of lung cancer.
The schematic diagrams of the atomizer used in combination with the wogonin inhalation solution are shown in figures 1 and 2.
The beneficial effects are that: the preparation method is simple through reasonable formula design, and the obtained liquid preparation has good stability and uniform particle size and meets the requirements of lung inhalation preparations. Can be used for treating acute lung injury, pulmonary fibrosis and lung cancer, and has definite therapeutic effect and better effect than positive medicine.
Drawings
Fig. 1 is a schematic diagram of a gas compression atomizer.
Fig. 2 is a schematic diagram of an atomizer for atomizing a liquid by means of a spring and a capillary tube.
Fig. 3 is an aerodynamic particle size distribution diagram of the aerosol inhalation solution 1.
FIG. 4 effect of inhalation wogonin on pathological changes of lung tissue in mice (pneumonia).
Figure 5 monitoring of aerosol stability during administration of wogonin pulmonary fibrosis for inhalation (pulmonary fibrosis).
FIG. 6 effect of inhalation wogonin on pathological changes of lung tissue in mice (pulmonary fibrosis).
FIG. 7 effect of inhalation wogonin on fibrosis-associated protein expression in mouse lung tissue (pulmonary fibrosis).
FIG. 8 effect of inhalation wogonin on collagen deposition in mouse lung tissue (pulmonary fibrosis).
FIG. 9 effect of inhalation wogonin on fluorescence intensity of lung cancer in situ (lung cancer) in mice at day 21.
Detailed Description
The technical solutions of the embodiments of the present invention will be clearly and completely described below in conjunction with the embodiments of the present invention, and it is apparent that the described embodiments are only some embodiments of the present invention, not all embodiments. All other embodiments, which can be made by those skilled in the art based on the embodiments of the invention without making any inventive effort, are intended to be within the scope of the invention.
The reagents and materials described in the examples are all commercially available. Wherein wogonin is provided by university of Chinese medical science.
Example 1
Preparation method of wogonin inhalation solution
The wogonin flavonoid compound has the conventional characteristics of the flavonoid compound: the water solubility is poor. The solubility of wogonin in conventional media and arginine media with different concentrations is studied in this example, and data support is provided for the preparation of wogonin inhalation solutions.
(1) About 50mg of wogonin (batch number: 20230226) was taken and put into 5 media of 10ml of 0.1M hydrochloric acid, 0.01M hydrochloric acid, pH=4.5 acetate buffer, pH=6.8 phosphate buffer and purified water, respectively, and the dissolution was observed by ultrasonic treatment for about 30 min. Filtering with 0.45 mu m PES filter membrane, and detecting content of the filtrate by HPLC method.
(2) Preparing 0.8%, 2.4% and 4% (m/v) arginine solution, taking about 50mg of the raw material medicine (batch number: 20230226), placing in 10ml of arginine water solution medium with different concentrations, and observing dissolution phenomenon by ultrasonic for 30 min. Filtering with 0.45 mu m PES filter membrane, and detecting content of the filtrate by HPLC method.
TABLE 1 solubility of wogonin in conventional Medium in 5
| Medium (D) | Saturated solubility μg/ml | pH value of |
| 0.1M hydrochloric acid | 0.33 | 1.03 |
| 0.01M hydrochloric acid | 0.30 | 1.95 |
| Acetate buffer at pH4.5 | 0.34 | 4.54 |
| phosphate buffer at pH6.8 | 0.89 | 6.86 |
| Purified water | 0.34 | 5.31 |
| 0.8% arginine solution | 1192.44 | 9.83 |
| 2.4% arginine solution | 4618.71 | 10.07 |
| 4% arginine solution | 7988.54 | 10.16 |
The above solubility study results showed that: the solubility of wogonin in 5 conventional mediums is low, and the arginine can obviously improve the solubility of wogonin.
In the present invention, we use arginine to increase the water solubility of wogonin to prepare inhalation solutions that can be used for nebulization.
The prescription is as follows:
table 2 wogonin inhalation solution prescription
| Composition of components | Aerosol inhalation solution 1 (23051201) |
| Wogonin | 10mg |
| Arginine (Arg) | 40mg |
| Water for injection | 2.5ml |
The preparation method comprises the following steps: arginine is dissolved in water for injection, and the prescription amount of raw material medicine is added for dissolving.
The aerodynamic particle size of the aerosol inhalation solution was analyzed using a cascade impactor and the results are shown in fig. 3.
Fig. 3 shows that: the median particle diameter of the inhalation solution in the aerosol state is 4.3 mu m, the proportion of inhalable fine particles (FPF%) is 57%, and the proportion can ensure that wogonin has higher lung deposition in the inhalation process, thereby being beneficial to fully playing the drug effect of wogonin.
Example 2
Preparation method of wogonin inhalation solution containing isotonic regulator
The embodiment provides a preparation method of wogonin inhalation solution containing an isotonic regulator, wherein the isotonic regulator is sodium chloride.
Table 3 prescription and preparation method of wogonin inhalation solution
The preparation method comprises the following steps: arginine and sodium chloride (if any in the prescription) are dissolved in water for injection, and the prescription amount of raw materials are added for dissolving.
In order to prevent the addition of the isotonic regulator sodium chloride from affecting wogonin, the invention compares whether the content of wogonin after the sodium chloride is added changes, and the result shows that: the addition of sodium chloride does not affect the wogonin content.
The results of the 0-day and 10-day contents are shown in Table 4.
TABLE 4 stability investigation of wogonin content with isotonic regulator
| Sample information | Determination of the concentration mg/ml | Compared with 0 day |
| 20230411-01-0h | 10.12 | 100.0％ |
| 20230411-01-24h | 9.91 | 97.9％ |
| 20230411-01-10 days | 10.05 | 99.3％ |
| 20230411-02-0h | 10.23 | 100.0％ |
| 20230411-02-24h | 9.93 | 97.1％ |
| 20230411-02-10 days | 10.00 | 97.8％ |
Example 3
Preparation method of wogonin inhalation solution capable of stably forming aerosol
In order to obtain an inhalation solution capable of generating stable and continuous aerosol, the embodiment provides a preparation method of wogonin inhalation solution, wherein the arginine dosage is 2% -10%. At the same time, citric acid can be used to adjust the pH of the inhalation solution if necessary.
Table 5 the wogonin inhalation solution formulation and method of preparation that can stably form an aerosol is as follows:
the preparation method comprises the following steps: arginine is dissolved in water for injection, and the prescription amount of raw material medicine is added for dissolving. Adjusting pH to 9-10 with citric acid.
After aerosol generation is carried out on the aerosol inhalation solution through an oral and nasal exposure system of a large animal, filter paper is adopted to collect the aerosol in different time periods, and the content is measured through HPLC. The aerosol concentrations for different concentrations and different time periods are shown in table 6. The results show that: the arginine dosage is within 2% -10%, and stable aerosol can be obtained. Indicating that the prescribed process results in an aerosol inhalation solution that produces a stable and consistent aerosol.
Table 6 drug delivery dose monitoring data in inhaled solution aerosols of different prescription compositions
Example 4
Research on main pharmacodynamics effect of wogonin for inhalation on LPS-induced lung injury of mice
1. Experimental materials
(1) Medicament
Example 1 wogonin solution for inhalation prepared in table 2.
(2) Experimental animal
C57BL/6J mice, 8 weeks old, male, were kept in SPF environment, were free to drink and were fed.
(3) Reagent(s)
1) Lipopolysaccharide (LPS): sigma Co., ltd., cargo number: l2880
2) PBS solution: naCl 8.0g, KH 2 PO 4 2.0g、Na 2 HPO 4 ·H 2 O1.56g and KCl 0.20g were dissolved in 1000mL of ultrapure water, sterilized by autoclaving, and stored at 4 ℃.
2. The experimental method comprises the following steps:
30 model mice are selected and randomly divided into 6 groups according to body weight, namely a blank control group, a model control group, a low-dose and high-dose test sample group and a positive drug control group, wherein each group comprises 6 mice. The grouping situation is shown in the following table:
TABLE 7 grouping of experimental animals
The model control group, the low-dose and high-dose sample-to-be-tested group and the positive drug control group mice are subjected to tracheal atomization injection of LPS (10 mg/kg), and a lung injury model of the mice is established. The sample group is subjected to intratracheal atomization administration after 3h of molding, 1 time per day for 3 days continuously, the administration volume is 0.1ml, the positive drug control group is subjected to intraperitoneal injection administration, 1 time per day, 3 days continuously, and the administration volume is 0.2ml.
The mice were weighed daily for a total of 4 times during the experiment.
On day 4 of the experiment, 3 mice per group were collected for routine blood testing. The remaining 3 mice in each group were bled and serum isolated and the levels of inflammatory factors IL-1. Beta., IL-6 and TNF-alpha were measured in the mouse serum by ELISA experiments.
All mice were euthanized after blood collection, lungs were collected and weighed for wet weight, lung tissue was lyophilized at-80 ℃, and lung dry weight was weighed and the lung wet weight to dry weight ratio was measured.
The method comprises the steps of taking a mouse alveolar lavage fluid, detecting the levels of inflammatory factors IL-1 beta, IL-6 and TNF-alpha in the lavage fluid through an ELISA (enzyme-linked immunosorbent assay), detecting the total protein concentration in the lavage fluid through a BCA (broadcast-specific area) experiment, and counting white blood cells of the lavage fluid.
Mice lungs were fixed with 4% paraformaldehyde and then sectioned in paraffin for HE staining.
3. Statistical methods:
all metering data toThe comparison between groups is shown using one-way variance (ANOVA) analysis. The difference was considered statistically significant with P < 0.05.
4. Experimental results:
the results of the study on LPS-induced lung injury symptoms of mice by inhalation wogonin show that compared with the model group, the inhalation wogonin (20, 40 mg/kg) can significantly reduce the number of white blood cells in alveolar lavage fluid (table 8), reduce the expression of inflammatory factors in mouse serum and alveolar lavage fluid (table 9), improve the wet weight-dry weight ratio of the lung of the mice (table 10), and improve the pathological indication of the lung tissue of the mice (fig. 4). From the above results, it was found that the inhalation wogonin solution has a good therapeutic effect on mice with acute lung injury induced by lipopolysaccharide.
TABLE 8 influence of wogonin for inhalation on the number of leukocytes in mouse alveolar lavage fluidn＝3)
* P < 0.05, P < 0.01 compared with the blank control group, # P＜0.05， ## p < 0.01 compared with model control group
TABLE 9 influence of wogonin for inhalation on cytokine expression in mouse alveolar lavage fluidn＝3)
* P < 0.05, P < 0.01 compared with the blank control group, # P＜0.05， ## p < 0.01 compared with model control group
TABLE 10 influence of wogonin for inhalation on wet weight to dry weight ratio of mouse lungsn＝3)
* P < 0.05, P < 0.01 compared with the blank control group, # P＜0.05， ## p < 0.01 compared with model control group
Example 5
Study of main pharmacodynamics effect of inhalation wogonin on bleomycin-induced pulmonary fibrosis of mice
1. Experimental materials
(1) Medicament
The wogonin solution for inhalation prepared in example 3.
(2) Experimental animal
C57BL/6J mice, 8 weeks old, male, were kept in SPF environment, were free to drink and were fed.
(3) Reagent(s)
1) Bleomycin (Bleomycin): MCE company, cat No.: HY-17565A.
2) PBS solution: naCl 8.0g, KH 2 PO 4 2.0g、Na 2 HPO 4 ·H 2 1.56g of O and 0.20g of KCl are dissolved in 1000mL of ultrapure water, sterilized by high-pressure steam and stored at 4 ℃.
2. The experimental method comprises the following steps:
the 40 model mice are randomly divided into 6 groups according to the body weight, namely a blank control group, a model control group, a low-dose and high-dose test sample group and a positive drug control group, and 8 mice in each group are respectively selected. The grouping situation is shown in the following table:
table 11 experimental animal groups
The model control group, the low-dose and high-dose sample of the tested sample and the positive drug control group are subjected to air tube atomization injection of bleomycin (5 mg/kg) to establish a pulmonary fibrosis model of the mice. The sample group was administered by oral and nasal exposure for the small animals for the first time 3 hours, 1 time per day, for 28 days continuously, the positive drug control group was administered by intraperitoneal injection 1 time per day, for 28 days continuously, and the administration volume was 0.2mL.
During the experiment, aerosol drug concentrations were continuously monitored during the dosing period in order to monitor drug stability during dosing.
The mice were weighed daily for a total of 4 times during the experiment.
Left mouse lung leaves were lavaged, fixed with 4% paraformaldehyde (first, 4% paraformaldehyde solution tissue was perfused through the bronchi) and paraffin sections were sectioned for HE staining.
On day 28 of the experiment, 3 mice per group were collected and serum was isolated and murine IL-1β, IL-6 and TNF- α levels were determined using the Elisa method.
On day 28 of the experiment, 3 mice were taken from each group to extract mRNA, and transcript levels of Acta2, col1a1 and Fn1 were detected by RT-qPCR.
On day 28 of the experiment, left lung leaves of mice were taken, lavaged, fixed with 4% paraformaldehyde (first, 4% paraformaldehyde solution tissue was perfused through bronchi), sectioned in paraffin, immunohistochemically stained, and examined for expression of fibrosis-related proteins.
On day 28 of the experiment, left lung lobes of mice were lavaged, fixed with 4% paraformaldehyde (first, 4% paraformaldehyde solution tissue was perfused through the bronchi) and paraffin sections were sectioned for sirius red staining and Masson staining.
3. Statistical methods:
all metering data toThe comparison between groups is shown using one-way variance (ANOVA) analysis. The difference was considered statistically significant with P < 0.05.
4. Experimental results:
the aerosol concentration was continuously monitored during the administration period as shown in fig. 5, and the results showed that the aerosol concentration was stable during the administration period. In the lung fibrosis model mice, the wogonin for inhalation can obviously inhibit the expression of inflammatory cytokines in serum of the lung fibrosis model mice induced by bleomycin (table 12), inhibit the transcription level of fibrosis related proteins in lung tissues of the mice (table 13), improve the pathological characteristics of the lung fibrosis (fig. 6), the expression of fibrosis related proteins in the lung tissues (fig. 7) and the influence of collagen deposition of the lung tissues of the mice (fig. 8), and the therapeutic effect of the medicament is dose-dependent and is superior to that of dexamethasone which is a positive medicament. In conclusion, the wogonin for inhalation has good treatment effect on the pulmonary fibrosis of mice induced by the bleomycin.
TABLE 12 influence of wogonin for inhalation on cytokine expression in mouse serumn＝3)
* P < 0.05, P < 0.01 compared with the blank control group, # P＜0.05， ## p < 0.01 compared with model control group
TABLE 13 influence of wogonin for inhalation on the transcript levels of fibrosis-related proteins in mouse lung tissuen＝3)
* P < 0.05, P < 0.01 compared with the blank control group, # P＜0.05， ## p < 0.01 compared with model control group
Example 6
Research on pharmacodynamics effect of wogonin for inhalation on in-situ lung cancer
1. Experimental materials
(1) Medicament
The wogonin solution for inhalation prepared in example 3.
(2) Experimental animal
BALB/c Nude mice, 18-22 g, 6 week old, male, 20.
2. The experimental method comprises the following steps:
the 12 model mice are selected and randomly divided into 2 groups according to the weight, namely a model control group and a tested sample group, and 6 mice are selected from each group. The grouping situation is shown in the following table:
table 14 experimental animal groups
Dosing volume: 100 μl of
Route of administration: tracheal atomization administration
Duration of administration: 21 days
Daily observations of the clinical status of animals, including tumor size measurements, mental status, mobility, diet, drinking water, etc., were made from inoculation to the end of the experiment. In vivo imaging was performed 2 times a week after tumor implantation, and fluorescence change curves were plotted.
3. Statistical methods:
the results of experiments on body weights and the like of the animals in each group are expressed as mean.+ -. Standard deviation (mean.+ -. SEM). Multiple group comparisons the presence or absence of significant differences between different treatment groups compared to the control group was compared using an analytical of variance (one way ANOVA/two way ANOVA) test method. Data were analyzed using Graphpad Prism. P < 0.05 is a significant difference.
4. Experimental results:
by day 21 of administration, the inhaled wogonin had a certain inhibition of tumor cell activity compared to the model group, with an inhibition rate of 74.2% (fig. 9). In conclusion, the wogonin for inhalation has good effect on lung cancer treatment.
Claims (10)
1. The wogonin inhalation preparation is characterized by comprising wogonin inhalation solution and an atomizer matched with the wogonin inhalation solution for use.
2. The wogonin inhalation formulation according to claim 1, wherein the wogonin inhalation solution has a median droplet size in the range of: 1-5 μm.
3. The wogonin inhalation formulation according to claim 2, wherein the wogonin inhalation solution has a mist droplet size of: d (D) 10 0.5-1.0 μm, D 50 1.5-5.0 μm, D 90 3.0-10 μm.
4. The wogonin inhalation formulation according to claim 1, wherein the wogonin inhalation solution has a wogonin content of 0.1-5.0% (w/v), sodium chloride content of 0.85-0.95% (w/v) and arginine or lysine or histidine or cysteine content of 2-10% (w/v); the pH value is 7.0-11.0.
5. The wogonin inhalation formulation of claim 1, wherein the nebuliser used in combination with the wogonin inhalation solution comprises a gas compression nebuliser, a nebuliser to effect nebulisation of a liquid by means of a spring and a capillary tube.
6. The method for preparing a wogonin inhalation formulation according to any one of claims 1 to 5, comprising the steps of: the preparation method comprises dissolving arginine in injectable water, adding wogonin, and ultrasound, or further heating or boiling to obtain the final product.
7. Use of a wogonin inhalation formulation according to claim 1 for the preparation of a medicament for the treatment of acute lung injury, pulmonary fibrosis or lung cancer.
8. The use according to claim 7, wherein the acute lung injury is 50 mg-300 mg/60 kg/person on a wogonin daily basis.
9. The use according to claim 7, wherein the pulmonary fibrosis is 200 mg-800 mg/60 kg/person on a wogonin daily basis.
10. The use according to claim 7, wherein lung cancer is 50 mg-300 mg/60 kg/person on a wogonin daily basis.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202311064407.6A CN117045624A (en) | 2023-08-22 | 2023-08-22 | Wogonin inhalation preparation and preparation method and medical application thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202311064407.6A CN117045624A (en) | 2023-08-22 | 2023-08-22 | Wogonin inhalation preparation and preparation method and medical application thereof |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN117045624A true CN117045624A (en) | 2023-11-14 |
Family
ID=88653244
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202311064407.6A Pending CN117045624A (en) | 2023-08-22 | 2023-08-22 | Wogonin inhalation preparation and preparation method and medical application thereof |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN117045624A (en) |
-
2023
- 2023-08-22 CN CN202311064407.6A patent/CN117045624A/en active Pending
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| JP2013136564A (en) | Method and device for provocation of air passage narrowing and/or induction of sputum | |
| EA003328B1 (en) | Pulmonary and nasal delivery of raloxifene | |
| WO2024017161A1 (en) | Fudosteine nebulization inhalation solution composition, drug assembly, and use thereof | |
| KR20210043601A (en) | Compositions containing mucolytic agents for treating mucus hypersecretion and devices for administration thereof | |
| CN117045624A (en) | Wogonin inhalation preparation and preparation method and medical application thereof | |
| CN114903889B (en) | Lizatriptan nasal spray and preparation method thereof | |
| US6462090B1 (en) | Formulations for detecting asthma | |
| CN114668788A (en) | Application of blumea oil inclusion compound in preparation of medical spray for treating asthma | |
| CN117100736A (en) | Application of compound QL-H008 in preparation of medicines for preventing and treating pulmonary diseases | |
| Solanki et al. | Dextran microparticulate inhalable dry powder for the treatment of cystic fibrosis and mucopolysaccharidosis | |
| CN107243024B (en) | A kind of double golden pharmaceutical compositions connected, a kind of Neulized inhalation solution of double gold even and its preparation method and application | |
| CN115068549B (en) | Atomized product for treating acute bronchitis cough of children and preparation method thereof | |
| Alaboud | In-vitro inhalation performance for formoterol dry powder and metred dose inhalers. In-vitro characteristics of the emitted dose from the formoterol dry powder and metred dose inhalers to identify the influence of inhalation flow, inhalation volume and the number of inhalation per dose. | |
| TWI790483B (en) | Care composition and use thereof | |
| CN111529491B (en) | Quick-acting safe ticagrelor oral spray and preparation method thereof | |
| WO2022077358A1 (en) | Care compositions and use thereof | |
| CN116983288A (en) | Application of wogonin preparation for oral and nasal inhalation in preparation of medicine for treating brain glioma | |
| CN113491676B (en) | Solution for atomizing inhalation of ribavirin and preparation method thereof | |
| RU2799050C2 (en) | Application of pulsatilla chinensis extract for obtaining a drug for the treatment of viral and/or bacterial diseases | |
| CN117159661A (en) | Traditional Chinese medicine composition, preparation and application thereof | |
| CN116327743A (en) | Application of baicalein inhalation preparation in preparation of acute lung injury treatment drugs | |
| CN115137713A (en) | Amphotericin B atomization inhalation preparation and preparation method thereof | |
| CN114425057A (en) | Care composition and use thereof | |
| CN110664819A (en) | Oleanolic acid preparation for inhalation, preparation method and medical application thereof | |
| CN117137971A (en) | Oviductus Ranae composition and Chinese medicinal composition for relieving cough and asthma, and preparation method and application thereof |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination |