CN114558136A - 碱性氨基酸修饰氨基四苯基卟啉化合物治疗骨髓炎的用途 - Google Patents
碱性氨基酸修饰氨基四苯基卟啉化合物治疗骨髓炎的用途 Download PDFInfo
- Publication number
- CN114558136A CN114558136A CN202210342791.0A CN202210342791A CN114558136A CN 114558136 A CN114558136 A CN 114558136A CN 202210342791 A CN202210342791 A CN 202210342791A CN 114558136 A CN114558136 A CN 114558136A
- Authority
- CN
- China
- Prior art keywords
- osteomyelitis
- group
- amino acid
- treatment
- pact
- 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
- 206010031252 Osteomyelitis Diseases 0.000 title claims abstract description 44
- -1 amino acid modified amino tetraphenylporphyrin compound Chemical class 0.000 title claims abstract description 35
- 239000003814 drug Substances 0.000 claims abstract description 26
- 238000002428 photodynamic therapy Methods 0.000 claims abstract description 9
- 238000001727 in vivo Methods 0.000 claims description 5
- 210000000988 bone and bone Anatomy 0.000 abstract description 35
- 208000015181 infectious disease Diseases 0.000 abstract description 25
- 241000283973 Oryctolagus cuniculus Species 0.000 abstract description 18
- 229940079593 drug Drugs 0.000 abstract description 17
- 241000894006 Bacteria Species 0.000 abstract description 15
- 241000191967 Staphylococcus aureus Species 0.000 abstract description 15
- 238000000034 method Methods 0.000 abstract description 12
- 150000001413 amino acids Chemical class 0.000 abstract description 8
- 239000003242 anti bacterial agent Substances 0.000 abstract description 5
- 230000002757 inflammatory effect Effects 0.000 abstract description 5
- 229940088710 antibiotic agent Drugs 0.000 abstract description 4
- 238000002474 experimental method Methods 0.000 abstract description 4
- 230000001404 mediated effect Effects 0.000 abstract description 4
- 230000008569 process Effects 0.000 abstract description 4
- 230000035876 healing Effects 0.000 abstract description 3
- 230000028327 secretion Effects 0.000 abstract description 3
- 230000002195 synergetic effect Effects 0.000 abstract description 2
- 238000002560 therapeutic procedure Methods 0.000 abstract 1
- 230000003115 biocidal effect Effects 0.000 description 54
- 235000001014 amino acid Nutrition 0.000 description 16
- 230000006378 damage Effects 0.000 description 15
- 210000002303 tibia Anatomy 0.000 description 15
- JLSFGHJZDAFBQP-UHFFFAOYSA-N 5,10,15,20-tetraphenyl-21,23-dihydroporphyrin-2-amine Chemical class NC1=C2NC(=C1)C(=C1C=CC(=N1)C(=C1C=CC(N1)=C(C=1C=CC(N=1)=C2C1=CC=CC=C1)C1=CC=CC=C1)C1=CC=CC=C1)C1=CC=CC=C1 JLSFGHJZDAFBQP-UHFFFAOYSA-N 0.000 description 14
- 230000000844 anti-bacterial effect Effects 0.000 description 13
- 238000000465 moulding Methods 0.000 description 13
- 230000000694 effects Effects 0.000 description 12
- YMWUJEATGCHHMB-UHFFFAOYSA-N Dichloromethane Chemical compound ClCCl YMWUJEATGCHHMB-UHFFFAOYSA-N 0.000 description 10
- 230000037396 body weight Effects 0.000 description 10
- 150000001875 compounds Chemical class 0.000 description 10
- CEAZRRDELHUEMR-URQXQFDESA-N Gentamicin Chemical compound O1[C@H](C(C)NC)CC[C@@H](N)[C@H]1O[C@H]1[C@H](O)[C@@H](O[C@@H]2[C@@H]([C@@H](NC)[C@@](C)(O)CO2)O)[C@H](N)C[C@@H]1N CEAZRRDELHUEMR-URQXQFDESA-N 0.000 description 9
- 208000027418 Wounds and injury Diseases 0.000 description 9
- 230000002829 reductive effect Effects 0.000 description 9
- 210000004872 soft tissue Anatomy 0.000 description 9
- 230000000007 visual effect Effects 0.000 description 9
- 108010048233 Procalcitonin Proteins 0.000 description 8
- 206010052428 Wound Diseases 0.000 description 8
- 230000001580 bacterial effect Effects 0.000 description 8
- 230000015572 biosynthetic process Effects 0.000 description 8
- CWCXERYKLSEGEZ-KDKHKZEGSA-N procalcitonin Chemical compound C([C@@H](C(=O)N1CCC[C@H]1C(=O)N[C@@H](CCC(N)=O)C(=O)N[C@H](C(=O)N[C@@H](C)C(=O)N[C@@H]([C@@H](C)CC)C(=O)NCC(=O)N[C@@H](C(C)C)C(=O)NCC(=O)N[C@@H](C)C(=O)N1[C@@H](CCC1)C(=O)NCC(O)=O)[C@@H](C)O)NC(=O)[C@@H](NC(=O)[C@H](CC=1NC=NC=1)NC(=O)[C@H](CC=1C=CC=CC=1)NC(=O)[C@H](CCCCN)NC(=O)[C@H](CC(N)=O)NC(=O)[C@H](CC=1C=CC=CC=1)NC(=O)[C@H](CC(O)=O)NC(=O)[C@H](CCC(N)=O)NC(=O)[C@@H](NC(=O)[C@H](CC=1C=CC(O)=CC=1)NC(=O)[C@@H](NC(=O)CNC(=O)[C@H](CC(C)C)NC(=O)[C@H](CCSC)NC(=O)[C@H]1NC(=O)[C@H]([C@@H](C)O)NC(=O)[C@H](CO)NC(=O)[C@H](CC(C)C)NC(=O)[C@H](CC(N)=O)NC(=O)CNC(=O)[C@@H](N)CSSC1)[C@@H](C)O)[C@@H](C)O)[C@@H](C)O)C1=CC=CC=C1 CWCXERYKLSEGEZ-KDKHKZEGSA-N 0.000 description 8
- 239000000243 solution Substances 0.000 description 8
- 210000001519 tissue Anatomy 0.000 description 8
- 229930182566 Gentamicin Natural products 0.000 description 7
- 206010061218 Inflammation Diseases 0.000 description 7
- 238000010171 animal model Methods 0.000 description 7
- 210000001185 bone marrow Anatomy 0.000 description 7
- 229960002518 gentamicin Drugs 0.000 description 7
- 230000004054 inflammatory process Effects 0.000 description 7
- 210000000056 organ Anatomy 0.000 description 7
- 125000002924 primary amino group Chemical group [H]N([H])* 0.000 description 7
- 230000008961 swelling Effects 0.000 description 7
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 description 6
- RJQXTJLFIWVMTO-TYNCELHUSA-N Methicillin Chemical compound COC1=CC=CC(OC)=C1C(=O)N[C@@H]1C(=O)N2[C@@H](C(O)=O)C(C)(C)S[C@@H]21 RJQXTJLFIWVMTO-TYNCELHUSA-N 0.000 description 6
- 230000036760 body temperature Effects 0.000 description 6
- 230000008859 change Effects 0.000 description 6
- 230000007423 decrease Effects 0.000 description 6
- 238000001514 detection method Methods 0.000 description 6
- 229960003085 meticillin Drugs 0.000 description 6
- QHGUCRYDKWKLMG-UHFFFAOYSA-N octopamine Chemical class NCC(O)C1=CC=C(O)C=C1 QHGUCRYDKWKLMG-UHFFFAOYSA-N 0.000 description 6
- 230000003247 decreasing effect Effects 0.000 description 5
- 230000002401 inhibitory effect Effects 0.000 description 5
- 230000001678 irradiating effect Effects 0.000 description 5
- 229960001576 octopamine Drugs 0.000 description 5
- 239000002244 precipitate Substances 0.000 description 5
- 231100000331 toxic Toxicity 0.000 description 5
- 230000002588 toxic effect Effects 0.000 description 5
- RTZKZFJDLAIYFH-UHFFFAOYSA-N Diethyl ether Chemical compound CCOCC RTZKZFJDLAIYFH-UHFFFAOYSA-N 0.000 description 4
- WSFSSNUMVMOOMR-UHFFFAOYSA-N Formaldehyde Chemical compound O=C WSFSSNUMVMOOMR-UHFFFAOYSA-N 0.000 description 4
- 241001465754 Metazoa Species 0.000 description 4
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 4
- 239000012984 antibiotic solution Substances 0.000 description 4
- 238000006243 chemical reaction Methods 0.000 description 4
- 230000007547 defect Effects 0.000 description 4
- 238000000338 in vitro Methods 0.000 description 4
- 210000000629 knee joint Anatomy 0.000 description 4
- 239000003504 photosensitizing agent Substances 0.000 description 4
- 238000004659 sterilization and disinfection Methods 0.000 description 4
- 239000000725 suspension Substances 0.000 description 4
- VHUUQVKOLVNVRT-UHFFFAOYSA-N Ammonium hydroxide Chemical compound [NH4+].[OH-] VHUUQVKOLVNVRT-UHFFFAOYSA-N 0.000 description 3
- 206010041925 Staphylococcal infections Diseases 0.000 description 3
- 208000033809 Suppuration Diseases 0.000 description 3
- ZMANZCXQSJIPKH-UHFFFAOYSA-N Triethylamine Chemical compound CCN(CC)CC ZMANZCXQSJIPKH-UHFFFAOYSA-N 0.000 description 3
- 206010048038 Wound infection Diseases 0.000 description 3
- 206010000269 abscess Diseases 0.000 description 3
- 235000011114 ammonium hydroxide Nutrition 0.000 description 3
- 210000000436 anus Anatomy 0.000 description 3
- 244000052616 bacterial pathogen Species 0.000 description 3
- 238000000576 coating method Methods 0.000 description 3
- 239000003085 diluting agent Substances 0.000 description 3
- 206010020718 hyperplasia Diseases 0.000 description 3
- 230000000415 inactivating effect Effects 0.000 description 3
- 210000002966 serum Anatomy 0.000 description 3
- 208000015339 staphylococcus aureus infection Diseases 0.000 description 3
- 230000001954 sterilising effect Effects 0.000 description 3
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 3
- QCBCPALLWXTPLW-SFHVURJKSA-N (2S)-2-(3,4-dihydroxyphenyl)-8,8-dimethyl-2,3,9,10-tetrahydropyrano[2,3-h]chromen-4-one Chemical compound C1([C@@H]2CC(=O)C=3C=CC4=C(C=3O2)CCC(O4)(C)C)=CC=C(O)C(O)=C1 QCBCPALLWXTPLW-SFHVURJKSA-N 0.000 description 2
- 206010059866 Drug resistance Diseases 0.000 description 2
- 206010015150 Erythema Diseases 0.000 description 2
- 206010028851 Necrosis Diseases 0.000 description 2
- 206010031264 Osteonecrosis Diseases 0.000 description 2
- DTQVDTLACAAQTR-UHFFFAOYSA-N Trifluoroacetic acid Chemical compound OC(=O)C(F)(F)F DTQVDTLACAAQTR-UHFFFAOYSA-N 0.000 description 2
- 230000001154 acute effect Effects 0.000 description 2
- 239000007853 buffer solution Substances 0.000 description 2
- 238000002512 chemotherapy Methods 0.000 description 2
- 239000011248 coating agent Substances 0.000 description 2
- 238000004440 column chromatography Methods 0.000 description 2
- 238000001804 debridement Methods 0.000 description 2
- 238000013461 design Methods 0.000 description 2
- 238000011161 development Methods 0.000 description 2
- 238000010790 dilution Methods 0.000 description 2
- 239000012895 dilution Substances 0.000 description 2
- 231100000321 erythema Toxicity 0.000 description 2
- 238000011156 evaluation Methods 0.000 description 2
- 239000001963 growth medium Substances 0.000 description 2
- 238000007490 hematoxylin and eosin (H&E) staining Methods 0.000 description 2
- 238000010562 histological examination Methods 0.000 description 2
- 210000004969 inflammatory cell Anatomy 0.000 description 2
- 238000002347 injection Methods 0.000 description 2
- 239000007924 injection Substances 0.000 description 2
- 239000007788 liquid Substances 0.000 description 2
- 210000003141 lower extremity Anatomy 0.000 description 2
- 244000005700 microbiome Species 0.000 description 2
- 239000000203 mixture Substances 0.000 description 2
- 230000004048 modification Effects 0.000 description 2
- 238000012986 modification Methods 0.000 description 2
- 230000017074 necrotic cell death Effects 0.000 description 2
- 230000007935 neutral effect Effects 0.000 description 2
- 238000011587 new zealand white rabbit Methods 0.000 description 2
- 210000000963 osteoblast Anatomy 0.000 description 2
- 239000012188 paraffin wax Substances 0.000 description 2
- WEXRUCMBJFQVBZ-UHFFFAOYSA-N pentobarbital Chemical compound CCCC(C)C1(CC)C(=O)NC(=O)NC1=O WEXRUCMBJFQVBZ-UHFFFAOYSA-N 0.000 description 2
- 239000003642 reactive oxygen metabolite Substances 0.000 description 2
- 230000009467 reduction Effects 0.000 description 2
- 239000011780 sodium chloride Substances 0.000 description 2
- 238000001228 spectrum Methods 0.000 description 2
- 238000007619 statistical method Methods 0.000 description 2
- 238000003756 stirring Methods 0.000 description 2
- 239000011550 stock solution Substances 0.000 description 2
- 230000008685 targeting Effects 0.000 description 2
- 230000008719 thickening Effects 0.000 description 2
- 231100000419 toxicity Toxicity 0.000 description 2
- 230000001988 toxicity Effects 0.000 description 2
- 210000003462 vein Anatomy 0.000 description 2
- 229920001817 Agar Polymers 0.000 description 1
- 206010002091 Anaesthesia Diseases 0.000 description 1
- 201000004569 Blindness Diseases 0.000 description 1
- 208000020084 Bone disease Diseases 0.000 description 1
- 206010005963 Bone formation increased Diseases 0.000 description 1
- 208000035473 Communicable disease Diseases 0.000 description 1
- 208000008960 Diabetic foot Diseases 0.000 description 1
- KCXVZYZYPLLWCC-UHFFFAOYSA-N EDTA Chemical compound OC(=O)CN(CC(O)=O)CCN(CC(O)=O)CC(O)=O KCXVZYZYPLLWCC-UHFFFAOYSA-N 0.000 description 1
- 238000002965 ELISA Methods 0.000 description 1
- 102000004190 Enzymes Human genes 0.000 description 1
- 108090000790 Enzymes Proteins 0.000 description 1
- 241000588724 Escherichia coli Species 0.000 description 1
- 241000192125 Firmicutes Species 0.000 description 1
- 206010023203 Joint destruction Diseases 0.000 description 1
- 206010024774 Localised infection Diseases 0.000 description 1
- FBVSXKMMQOZUNU-NSHDSACASA-N N2,N6-Bis{[(2-methyl-2-propanyl)oxy]carbonyl}lysine Chemical compound CC(C)(C)OC(=O)NCCCC[C@@H](C(O)=O)NC(=O)OC(C)(C)C FBVSXKMMQOZUNU-NSHDSACASA-N 0.000 description 1
- 208000002565 Open Fractures Diseases 0.000 description 1
- 206010031256 Osteomyelitis chronic Diseases 0.000 description 1
- 229930182555 Penicillin Natural products 0.000 description 1
- JGSARLDLIJGVTE-MBNYWOFBSA-N Penicillin G Chemical compound N([C@H]1[C@H]2SC([C@@H](N2C1=O)C(O)=O)(C)C)C(=O)CC1=CC=CC=C1 JGSARLDLIJGVTE-MBNYWOFBSA-N 0.000 description 1
- 208000006735 Periostitis Diseases 0.000 description 1
- 241000589517 Pseudomonas aeruginosa Species 0.000 description 1
- 241000700159 Rattus Species 0.000 description 1
- 208000034189 Sclerosis Diseases 0.000 description 1
- 238000010521 absorption reaction Methods 0.000 description 1
- 230000009471 action Effects 0.000 description 1
- 208000038016 acute inflammation Diseases 0.000 description 1
- 230000006022 acute inflammation Effects 0.000 description 1
- 230000000996 additive effect Effects 0.000 description 1
- 239000008272 agar Substances 0.000 description 1
- QGZKDVFQNNGYKY-UHFFFAOYSA-N ammonia Natural products N QGZKDVFQNNGYKY-UHFFFAOYSA-N 0.000 description 1
- 238000002266 amputation Methods 0.000 description 1
- 230000037005 anaesthesia Effects 0.000 description 1
- 238000000540 analysis of variance Methods 0.000 description 1
- 238000004458 analytical method Methods 0.000 description 1
- 210000001557 animal structure Anatomy 0.000 description 1
- 230000003042 antagnostic effect Effects 0.000 description 1
- 230000002924 anti-infective effect Effects 0.000 description 1
- 230000000845 anti-microbial effect Effects 0.000 description 1
- 239000004599 antimicrobial Substances 0.000 description 1
- 238000013459 approach Methods 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 230000008901 benefit Effects 0.000 description 1
- 210000004369 blood Anatomy 0.000 description 1
- 239000008280 blood Substances 0.000 description 1
- 238000010241 blood sampling Methods 0.000 description 1
- 230000004579 body weight change Effects 0.000 description 1
- 210000002449 bone cell Anatomy 0.000 description 1
- 201000011510 cancer Diseases 0.000 description 1
- 230000001413 cellular effect Effects 0.000 description 1
- 230000001684 chronic effect Effects 0.000 description 1
- 208000037976 chronic inflammation Diseases 0.000 description 1
- 230000006020 chronic inflammation Effects 0.000 description 1
- 230000002301 combined effect Effects 0.000 description 1
- 230000034994 death Effects 0.000 description 1
- 206010012601 diabetes mellitus Diseases 0.000 description 1
- 238000007865 diluting Methods 0.000 description 1
- 201000010099 disease Diseases 0.000 description 1
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 1
- GRIXGZQULWMCLU-UHFFFAOYSA-L disodium;7-[[2-carboxylato-2-(4-hydroxyphenyl)acetyl]amino]-7-methoxy-3-[(1-methyltetrazol-5-yl)sulfanylmethyl]-8-oxo-5-oxa-1-azabicyclo[4.2.0]oct-2-ene-2-carboxylate Chemical compound [Na+].[Na+].C12OCC(CSC=3N(N=NN=3)C)=C(C([O-])=O)N2C(=O)C1(OC)NC(=O)C(C([O-])=O)C1=CC=C(O)C=C1 GRIXGZQULWMCLU-UHFFFAOYSA-L 0.000 description 1
- 239000012153 distilled water Substances 0.000 description 1
- 238000005553 drilling Methods 0.000 description 1
- 230000000816 effect on animals Effects 0.000 description 1
- 239000003480 eluent Substances 0.000 description 1
- 210000002745 epiphysis Anatomy 0.000 description 1
- RIFGWPKJUGCATF-UHFFFAOYSA-N ethyl chloroformate Chemical compound CCOC(Cl)=O RIFGWPKJUGCATF-UHFFFAOYSA-N 0.000 description 1
- 230000003203 everyday effect Effects 0.000 description 1
- 210000003414 extremity Anatomy 0.000 description 1
- 238000000556 factor analysis Methods 0.000 description 1
- 210000003195 fascia Anatomy 0.000 description 1
- 239000000706 filtrate Substances 0.000 description 1
- 238000001914 filtration Methods 0.000 description 1
- 230000037406 food intake Effects 0.000 description 1
- 235000012631 food intake Nutrition 0.000 description 1
- 230000008014 freezing Effects 0.000 description 1
- 238000007710 freezing Methods 0.000 description 1
- 238000000227 grinding Methods 0.000 description 1
- 210000002216 heart Anatomy 0.000 description 1
- 230000002390 hyperplastic effect Effects 0.000 description 1
- 239000005457 ice water Substances 0.000 description 1
- 239000007943 implant Substances 0.000 description 1
- 238000011534 incubation Methods 0.000 description 1
- 208000014674 injury Diseases 0.000 description 1
- 230000003993 interaction Effects 0.000 description 1
- 230000001788 irregular Effects 0.000 description 1
- 210000003734 kidney Anatomy 0.000 description 1
- 230000002147 killing effect Effects 0.000 description 1
- 238000002372 labelling Methods 0.000 description 1
- 210000004185 liver Anatomy 0.000 description 1
- 231100000053 low toxicity Toxicity 0.000 description 1
- 210000004072 lung Anatomy 0.000 description 1
- 235000018977 lysine Nutrition 0.000 description 1
- 150000002669 lysines Chemical class 0.000 description 1
- 238000005259 measurement Methods 0.000 description 1
- 210000003205 muscle Anatomy 0.000 description 1
- 239000013642 negative control Substances 0.000 description 1
- 230000000399 orthopedic effect Effects 0.000 description 1
- 230000011164 ossification Effects 0.000 description 1
- 206010033675 panniculitis Diseases 0.000 description 1
- 230000001575 pathological effect Effects 0.000 description 1
- 229940049954 penicillin Drugs 0.000 description 1
- 229960001412 pentobarbital Drugs 0.000 description 1
- 210000003460 periosteum Anatomy 0.000 description 1
- 230000002980 postoperative effect Effects 0.000 description 1
- 230000001376 precipitating effect Effects 0.000 description 1
- 238000002360 preparation method Methods 0.000 description 1
- 238000002203 pretreatment Methods 0.000 description 1
- 238000012545 processing Methods 0.000 description 1
- 239000000047 product Substances 0.000 description 1
- 230000000750 progressive effect Effects 0.000 description 1
- 238000004445 quantitative analysis Methods 0.000 description 1
- 238000011084 recovery Methods 0.000 description 1
- 230000004044 response Effects 0.000 description 1
- 238000005070 sampling Methods 0.000 description 1
- 229960005491 sodium morrhuate Drugs 0.000 description 1
- 239000007787 solid Substances 0.000 description 1
- 239000002904 solvent Substances 0.000 description 1
- 210000000952 spleen Anatomy 0.000 description 1
- 238000010186 staining Methods 0.000 description 1
- 210000004304 subcutaneous tissue Anatomy 0.000 description 1
- 239000013589 supplement Substances 0.000 description 1
- 238000003786 synthesis reaction Methods 0.000 description 1
- 230000009885 systemic effect Effects 0.000 description 1
- 238000012360 testing method Methods 0.000 description 1
- 230000001225 therapeutic effect Effects 0.000 description 1
- 229910021642 ultra pure water Inorganic materials 0.000 description 1
- 239000012498 ultrapure water Substances 0.000 description 1
- 238000010200 validation analysis Methods 0.000 description 1
- 230000035899 viability Effects 0.000 description 1
- 238000005406 washing Methods 0.000 description 1
- 239000001993 wax Substances 0.000 description 1
- 230000003442 weekly effect Effects 0.000 description 1
- 238000005303 weighing Methods 0.000 description 1
- 230000029663 wound healing Effects 0.000 description 1
Images
Classifications
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K41/00—Medicinal preparations obtained by treating materials with wave energy or particle radiation ; Therapies using these preparations
- A61K41/0057—Photodynamic therapy with a photosensitizer, i.e. agent able to produce reactive oxygen species upon exposure to light or radiation, e.g. UV or visible light; photocleavage of nucleic acids with an agent
- A61K41/0071—PDT with porphyrins having exactly 20 ring atoms, i.e. based on the non-expanded tetrapyrrolic ring system, e.g. bacteriochlorin, chlorin-e6, or phthalocyanines
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P19/00—Drugs for skeletal disorders
- A61P19/08—Drugs for skeletal disorders for bone diseases, e.g. rachitism, Paget's disease
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
- A61P31/04—Antibacterial agents
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P7/00—Drugs for disorders of the blood or the extracellular fluid
-
- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
- Y02A50/00—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE in human health protection, e.g. against extreme weather
- Y02A50/30—Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change
Landscapes
- Health & Medical Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- General Health & Medical Sciences (AREA)
- Veterinary Medicine (AREA)
- Public Health (AREA)
- Medicinal Chemistry (AREA)
- Animal Behavior & Ethology (AREA)
- Pharmacology & Pharmacy (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- General Chemical & Material Sciences (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Organic Chemistry (AREA)
- Physical Education & Sports Medicine (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Engineering & Computer Science (AREA)
- Rheumatology (AREA)
- Orthopedic Medicine & Surgery (AREA)
- Communicable Diseases (AREA)
- Oncology (AREA)
- Biochemistry (AREA)
- Molecular Biology (AREA)
- Epidemiology (AREA)
- Diabetes (AREA)
- Hematology (AREA)
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
Abstract
本发明公开了碱性氨基酸修饰氨基四苯基卟啉化合物治疗骨髓炎的用途。所述的氨基酸修饰氨基四苯基卟啉化合物具有下述结构：
Description
技术领域
本发明属于药物领域，具体涉及碱性氨基酸修饰氨基四苯基卟啉化合物治疗骨髓炎的用途，特别是涉及四个赖氨酸修饰的氨基四苯基卟啉化合物在制备光动力治疗骨髓炎药物中的应用。
背景技术
骨髓炎是一种由感染微生物引起的炎症性骨疾病，能导致骨组织的进行性破坏和坏死[1]。骨髓炎可分为三种亚型：急性、亚急性和慢性。急性和亚急性骨髓炎主要发生在儿童和老年人中[2]。慢性骨髓炎在开放性骨折、糖尿病和耐药菌所致骨感染中较为常见[3]，约20％糖尿病足患者会并发骨髓炎[4]。骨髓炎能够导致骨缺损、骨坏死，具有较高的复发率。
目前骨髓炎的治疗方法主要包括手术清创和抗生素治疗[5]。手术清创很难彻底清除髓腔内残留的致病菌[6]。抗生素的广泛大剂量应用也会导致多重耐药菌株的产生，使得耐药菌株所致骨髓炎的治疗越来越棘手[7]。目前，骨髓炎因治疗过程缓慢，治愈率较低，病情严重的患者还会面临截肢的风险，成为骨科医生面临的难题之一[8]。因此，探寻一种具有疗效好、副作用少、能有效减少复发的治疗方法是临床医生迫切解决的问题。光动力抗菌化学疗法因毒副作用小，不易产生耐药性和抗菌谱广等优点而逐渐引起人们的关注[9]。
光动力抗菌化学疗法(Photodynamic Antimicrobial Chemotherapy，PACT)的原理是无毒副作用的光敏剂先特异性的聚集在细菌体内，然后用适当波长的可见光照射细菌，激活细菌体内的光敏剂以产生对细菌具有毒性的活性氧(ROS)，达到灭活细菌的目的[10]。近年来，其临床适应证已从恶性肿瘤扩展到越来越多的良性疾病。PACT被认为是一种能有效灭活多种微生物的方法，对于耐药菌及细菌形成的生物膜均有很好的灭菌效果。因其具有靶向性强、毒副作用低、抗菌谱广、起效快、可重复治疗和无耐药的特点，目前已在浅表局限性感染的治疗中被广泛应用。体外[11,12]和体内[13,14]的研究表明，PACT具有抗感染特性和促进伤口愈合的作用。目前，PACT在治疗浅表感染的报道相对较多，而PACT治疗骨髓炎鲜有文献报道。
本实验室设计合成了一系列具有良好理化性质的碱性氨基酸修饰氨基四苯基卟啉化合物，其中化合物LD4具有良好的水溶性、低毒和靶向性，在感染性疾病的治疗中表现出更广谱的杀菌效果[15]、更好的生物相容性及稳定性[16,17]。它能有效杀灭革兰氏阳性菌及革兰氏阴性菌，抑制炎症因子的分泌，促进伤口感染后创面愈合。前期实验表明，LD4对耐甲氧西林金黄色葡萄球菌(MRSA)、大肠杆菌和铜绿假单胞菌的体外灭菌效果良好；体内实验显示，LD4介导的PACT对三种混合细菌所致的大鼠创面感染的治疗效果也很好。
在骨髓炎的治疗中，全身使用抗生素会导致感染部位抗生素浓度较低，难以达到彻底灭活感染灶内致病菌的目的[18]。而PACT作为抗生素治疗的有益补充，可直接作用于骨感染区域，达到灭菌、清洁的治疗效果。本申请利用LD4介导的PACT治疗骨髓炎髓腔内、外的感染，PACT直接作用于感染区域，灭活感染灶内的致病菌，达到治疗骨髓炎的目的。
参考文献
[1]Daniel P L,Francis A W.Osteomyelitis[J].Lancet,2004,364(9431):369-379.
[2]Nicola K,Emily J R,Amro W,et al.Staphylococcal osteomyelitis:Diseaseprogression,treatment challenges,and future directions[J].2018,31(2):e00084-17.
[3]Maria D,Andrew J H,Jamie F,et al.The microbiology of chronicosteomyelitis:Changes over ten years[J].J Infect,2019,79(3):189-198.
[4]Terese G,Guido L.Current health and economic burden of chronicdiabeticosteomyelitis[J].Expert Rev Pharmacoecon Outcomes Res,2019,19(3):279-286.
[5]Takahiro N,Sang Y L,Takashi I,et al.Antibiotic-impregnated calciumphosphatecement as part of a comprehensive treatment for patients withestablished orthopaedic infection[J].JOrthop Sci,2016,21(4):539-545.
[6]Carmen C S,Cristin C,Florica B,et al.The influence of foreign bodysurface areaon the outcome of chronic osteomyelitis[J].Med Eng Phys,2016,38(9):870-876.
[7]Sheehy S H,Atkins B A,Bejon P,et al.The microbiology of chronicosteomyelitis:prevalence of resistance to common empirical anti-microbialregimens[J].J Infect,2010,60(5):338-343.
[8]Tschudin-Sutter S,Frei R,Dangel M,et al.Validation of a treatmentalgorithm fororthopaedic implant-related infections with device-retention-results from a prospectiveobservational cohort study[J].Clin MicrobiolInfect,2016,22(5):457.
[9]Anabela T,Carla M B,Maria A F,et al.Antimicrobial photodynamictherapy:studyof bacterial recovery viability and potential development ofresistance after treatment[J].Mar Drugs,2010,8(1):91-105.
[10]Wainwright M.Photodynamic antimicrobial chemotherapy(PACT)[J].JAntimicrob Chemother,1998,42(1):13-28.
[11]Sung S C,Hui Y O,Eui J K,et al.In vitro bactericidal effects ofphotodynamictherapy combined with four tetracyclines against clostridioidesdifficile KCTC5009 in planktoniccultures[J].Pathogens,2020,9(4):279.
[12]Heba E E,Tareq Y,Mona B M,et al.Photothermal versusphotodynamictreatment for the inactivation of the bacteria Escherichia coliand Bacillus cereus:An in vitrostudy[J].Photodiagnosis Photodyn Ther,2019,27:317-326.
[13]Liu X L,Chen R H,Lv J,et al.High-resolution bimodal imaging andpotentantibiotic/photodynamic synergistic therapy for osteomyelitis with abacterial inflammation-specificversatile agent[J].Acta Biomater,2019,99:363-372.
[14]Mai B J,Gao Y R,Li M,et al.Photodynamic antimicrobialchemotherapy forStaphylococcus aureus and multidrug-resistant bacterial burninfection in vitro and in vivo[J].Int JNanomedicine,2017,12:5915-5931.
[15]Meng S,Xu Z P,Hong G,et al.Synthesis,characterization and invitrophotodynamic antimicrobial activity of basic amino acid-porphyrinconjugates[J].Eur J Med Chem,2015,92:35-48.
[16]Zhao Z J,Ma J D,Wang Y Y,et al.Antimicrobial photodynamic therapycombinedwith antibiotic in the treatment of rats with third-degree burns[J].Front Microbiol,2021,12:622410.
[17]Xu Z P,Gao Y X,Meng S,et al.Mechanism and in vivo evaluation:photodynamicantibacterial chemotherapy of lysine-porphyrin conjugate[J].FrontMicrobiol,2016,7:242.
[18]Nalini R,Bruce H Z,Benjamin A L.Treating osteomyelitis:antibiotics andsurgery[J].Plast Reconstr Surg,2011,127(Suppl 1):177S-187S.
发明内容
本发明的目的在于克服现有技术的不足，提供碱性氨基酸修饰氨基四苯基卟啉化合物作为光动力治疗骨髓炎药物的用途。
本发明的目的主要通过以下技术方案实现：
碱性氨基酸修饰氨基四苯基卟啉化合物在制备光动力治疗骨髓炎药物的应用，所述的碱性氨基酸修饰氨基四苯基卟啉化合物具有下述结构：
所述的药物为光动力治疗药物。
实验证明：碱性氨基酸修饰氨基四苯基卟啉化合物在光动力治疗耐药菌感染所致的兔胫骨骨髓炎过程中，可以杀灭导致感染的金黄色葡萄球菌，抑制炎症因子的分泌，促进骨组织的愈合。同时，碱性氨基酸修饰氨基四苯基卟啉化合物介导的光动力疗法联合抗生素在骨髓炎治疗方面具有一定的协同作用，能够在一定程度上减少抗生素的使用剂量，为骨髓炎的临床治疗提供了新方法。
附图说明
图1为本发明实施例1的碱性氨基酸修饰氨基四苯基卟啉化合物LD4治疗耐药金黄色葡萄球菌感染所致的兔胫骨骨髓炎模型动物的体温和体重变化情况。
图2为本发明实施例1的碱性氨基酸修饰氨基四苯基卟啉化合物LD4治疗耐药金黄色葡萄球菌感染所致的兔胫骨骨髓炎模型动物胫骨组织的肉眼观察和X线检测结果。
图3为本发明实施例1的碱性氨基酸修饰氨基四苯基卟啉化合物LD4治疗耐药金黄色葡萄球菌感染所致的兔胫骨骨髓炎模型动物胫骨骨髓组织的细菌培养结果。
图4为本发明实施例1的碱性氨基酸修饰氨基四苯基卟啉化合物LD4治疗耐药金黄色葡萄球菌感染所致的兔胫骨骨髓炎模型动物胫骨的组织学检查结果。
图5为本发明实施例1的碱性氨基酸修饰氨基四苯基卟啉化合物LD4治疗耐药金黄色葡萄球菌感染所致的兔胫骨骨髓炎模型动物的血清降钙素原检测结果。
图6为本发明实施例1的碱性氨基酸修饰氨基四苯基卟啉化合物LD4治疗耐药金黄色葡萄球菌感染所致的兔胫骨骨髓炎模型动物各脏器组织的HE染色图。
具体实施方式
下面通过实施例对本发明作进一步说明，其目的仅在于更好的理解本发明的内容而非限制本发明的保护范围：
实施例1碱性氨基酸修饰氨基四苯基卟啉化合物(LD4)的合成
将Boc-Lys(Boc)-OH(467.67mg，1.35mmol)置于反应瓶中，N2保护下加入干燥的THF 20ml，磁子搅拌。冷却至-17℃，加入三乙胺(197.60μl，1.42mmol)和氯甲酸乙酯(131.10μl，1.38mmol)反应1h，生成白色沉淀，过滤，弃去沉淀。取四氨基四苯基卟啉(202.40mg，0.30mmol)溶解于15ml THF中，将上述滤液加入，室温搅拌反应14h。TLC(二氯甲烷：甲醇：氨水＝60:1:0.6)监测反应进程。反应完全后，将反应液倒入冰水中，析出沉淀，过滤，水洗3次，得到紫色固体。最后柱层析分离(洗脱剂：二氯甲烷：甲醇：氨水＝30:1:0.4)得产物596.13mg，产率93％。
将上述步骤得到的100.00mg样品用10ml干燥的CH2Cl2溶解，缓慢滴加三氟乙酸10ml，室温反应30min。旋除溶剂，加入无水乙醚，生成浅绿色沉淀，过滤，30ml二氯甲烷和30ml无水乙醚各洗涤3次。再用20ml蒸馏水将绿色沉淀溶解，氨水调节PH 7-8，析出紫色沉淀，过滤，水洗2次，柱层析分离得到碱性氨基酸修饰氨基四苯基卟啉化合物LD451.96mg，产率87％。
实施例2碱性氨基酸修饰氨基四苯基卟啉化合物(LD4)的体外光动力抗菌效果
本发明实施例1制备的碱性氨基酸修饰氨基四苯基卟啉化合物LD4最低抑菌浓度和最小杀菌浓度的测定，包括如下步骤：
实验菌株：耐甲氧西林金黄色葡萄球菌(MRSA)菌株由河北大学附属医院检验科临床分离提取，对青霉素类抗生素不敏感，对庆大霉素敏感。
将接种耐甲氧西林金黄色葡萄球菌的琼脂平板打开，用1μl灭菌接种环挑取边缘光滑完整的菌落，接种于5ml LB培养基中，置于37℃、200转/分的恒温震荡培养箱中，隔夜培养18小时。在9.5ml培养基中加入0.5ml菌液，每隔一小时取样一次，并用酶标仪测量OD值，通过稀释涂布平板法计算出相应OD值所对应的活菌菌落浓度。计算菌落浓度和OD值的对应线性关系，当OD600nm＝0.5时，所对应的菌液浓度为1.5×108CFU/ml。取上述浓度的耐甲氧西林金黄色葡萄球菌悬液备用。
96孔板每孔加入20μl的耐甲氧西林金黄色葡萄球菌混悬液(1.5×108CFU/ml)和180μl的LD4化合物(浓度分别为2、4、8、16、32、64、128、256和512μM)。37℃避光孵育30min，然后置于激光(650nm，2A)下照射30min，阴性对照不照光。照光后，在37℃培养箱中孵育18小时，然后评估碱性氨基酸修饰氨基四苯基卟啉化合物LD4的最低抑菌浓度和最小杀菌浓度。
LD4使细菌悬液从混浊到清澈明显变化所需的浓度称为最低抑菌浓度(MIC)，而平板上观察到5个或更少菌落的浓度称为最小杀菌浓度(MBC)。如表1所示，LD4具有很强的细菌光灭活能力，对MRSA的MIC值为4.0μM，MBC值为8.0μM。同时，LD4的暗毒性较低，MIC值和MBC值均大于500μM。从MBC值可以看出，LD4对MRSA有很强的杀菌活性。因此，LD4是一种较好的治疗耐药金黄色葡萄球菌所致骨髓炎的药物。
表1LD4对耐甲氧西林金黄色葡萄球菌的最低抑菌浓度(MIC，μM)和最小杀菌浓度(MBC，μM)
实施例3碱性氨基酸修饰氨基四苯基卟啉化合物(LD4)的体外光动力抗菌效果
本发明实施例1制备的碱性氨基酸修饰氨基四苯基卟啉化合物LD4与抗生素联用分数抑菌浓度指数(FICI)的测定，包括如下步骤：
用超纯水溶解庆大霉素注射液和LD4，得到浓度为4、2、1、0.5和0.25MIC的原液，以确定其分数抑菌浓度指数(FICI)。采用棋盘设计法，在96孔板的水平柱和垂直柱中分别加入不同浓度的50μl的每种药物。然后加入50μl细菌悬液(108CFU/ml)，轻轻摇晃混合。37℃避光孵育30min，然后置于激光(650nm，2A)下照射30min，照光后，在37℃培养箱中孵育18小时。对于每个抗菌药物组合，我们通过计算组合的MIC除以每个药物单独使用的MIC的比率，然后将这两个比率相加计算FICI：
FICI＝[MICA(withB)/MICA(alone)]+[MICB(withA)/MICB(alone)]
FICI的数据判定标准：协同性，FICI≤0.5；无相互作用，FICI 0.5-2.0；拮抗性，FICI>2.0。
庆大霉素与LD4联合应用于MRSA的平均FICI为0.625，表明这种组合存在相加效应。
表2庆大霉素注射液和LD4的联合应用效果
MICA：庆大霉素；MICB：碱性氨基酸修饰的氨基四苯基卟啉化合物LD4；MICA(withB)：庆大霉素与碱性氨基酸修饰的氨基四苯基卟啉化合物LD4；MICB(withA)：碱性氨基酸修饰的氨基四苯基卟啉化合物LD4与庆大霉素。
实施例4碱性氨基酸修饰氨基四苯基卟啉化合物(LD4)治疗骨髓炎
本发明实施例1制备的碱性氨基酸修饰氨基四苯基卟啉化合物LD4治疗耐药金黄色葡萄球菌感染所致的兔胫骨骨髓炎的在体实验过程，包括如下步骤：
40只新西兰大白兔称重后耳缘静脉注射3％戊巴比妥溶液(30mg/kg)，麻醉后将兔子头部后仰，四肢外展仰卧位固定，双侧后肢膝关节远端备皮至膝关节远端5cm处，碘伏消毒后铺无菌单。胫骨上段前内侧膝关节间隙下1cm为切口起点，沿胫骨方向做纵行切口，切开皮肤，逐层分离皮下、筋膜、肌肉，剥离骨膜，显露胫骨上段前内侧，用1mm牙科钻钻至髓腔，5ml注射器抽取骨髓约0.2ml，再用1ml注射器向髓腔内注射0.1ml鱼肝油酸钠溶液，5分钟后，用1ml注射器向髓腔内注射0.1ml的金黄色葡萄球菌溶液(1×108CFU/ml)，孔洞用无菌骨蜡封闭，逐层缝合。同法再对侧后肢进行造模。造模一周后随机处死4只兔子，取骨髓组织进行细菌培养，结果显示所有的兔子骨髓腔均有金葡菌存在，证实造模成功。
取造模成功的36只新西兰大白兔，随机分为4组：(a)模型对照组，(b)抗生素组，(c)PACT治疗组，(d)PACT+抗生素治疗组。PACT+抗生素组：伤口局部注射0.2ml的LD4溶液(40μM)，肌肉注射0.4ml的抗生素溶液，暗室孵育30分钟后进行激光照射，照射时间为15分钟，第二天不注射光敏剂，肌肉注射0.4ml的抗生素溶液，同样进行激光照射，照射时间为15分钟，此为一个疗程；PACT组除不注射抗生素溶液外，其余步骤与PACT+抗生素相同；抗生素组：伤局部注射0.2ml的生理盐水，每天肌肉注射0.4ml的抗生素溶液；对照组：伤口局部注射0.2ml的生理盐水，肌肉注射0.4ml的生理盐水。重复治疗7个疗程，治疗时间持续2周。
体温体重检测：术前及术后每周测量大白兔的体重，计算体重变化。术前及术后每天上午9：00用电子体温计测量兔子肛门温度，插入肛门深度2.0-3.0cm，待体温计发出“嘀嘀嘀”声之后再测30s，记录读数。
从造模前一天开始每天测量各组实验动物的体温。造模后5天内，各组体温均有明显的升高，但各组之间的变化无明显的差异(p>0.05)(图1，A)。与治疗前相比，治疗后各治疗组的体温均有明显的减低并逐渐趋于正常体温。PACT+抗生素组、PACT组和抗生素组的体温与对照组相比均有不同程度的下降，差异有统计学意义(p<0.05)，但各治疗组之间的体温变化无明显的差异(p>0.05)。
造模前测量实验动物的体重，造模后每周测量体重并进行统计分析(图1，B、C)。造模后各组体重与造模之前相比均有明显下降(p<0.01)；开始治疗后，各组体重与治疗前相比均有升高，其中以PACT+抗生素组和PACT组体重升高较为明显(p<0.05)，抗生素组体重升高较为缓慢(p<0.05)。与对照组相比，PACT+抗生素组、PACT组和抗生素组的体重变化有统计学意义(p<0.05)，其中以PACT+抗生素组为最显著，PACT组次之。
肉眼观察：术后观察大白兔的活动、进食等一般情况，记录大白兔切口部位的渗出情况、有无窦道形成及无脓液流出等，记录死亡情况。分别在治疗前、治疗2周和治疗4周后随机取出并处死实验动物，沿原切口进入暴露胫骨，观察骨形态及周围软组织情况。采用唐辉的方法，双盲完成肉眼观察评分：0分，无感染征象；1分，皮肤表面只有少量红斑；2分，皮肤表面有红斑且合并胫骨干增粗及窦道，或合并脓液渗出；3分，严重骨质吸收、脓肿。评分≧2分定义为明显感染。
肉眼观察能够直观的评价骨髓炎感染的严重程度及治疗效果。造模一周后，各组均有明显的周围软组织肿胀及伤口流脓，部分还有窦道的形成(图2，A)。在治疗2周后，对照组可见周围软组织的肿胀、窦道形成及骨质的破坏。在治疗5周后，对照组感染程度仍在继续加重，有明显的骨质破坏及死骨形成。对照组的软组织肿胀、窦道形成及骨质破坏程度均较治疗前明显，感染程度随着时间的增加而加重。抗生素组在治疗2周后，感染程度较前未见明显变化，但感染程度明显较对照组轻(p<0.05)。抗生素组在治疗5周后，感染程度较治疗前明显加重，但与对照组之间的差别不大(p>0.05)。PACT组和PACT+抗生素组在2周后，伤口感染及骨破坏程度与对照组相比均有减轻，其中以PACT+抗生素组减轻最为明显(图2，A、B)。PACT+抗生素组在治疗5周后未见明显的骨质破坏，骨髓炎基本趋于愈合。
通过对软组织肿胀、脓肿、窦道形成及骨质破坏进行定量分析。各组的肉眼评分在治疗前未见明显的差异(p>0.05)。随着时间的增加，对照组肉眼评分均较治疗前有明显的升高，各治疗组的肉眼评分与对照组相比均有减低，其中PACT+抗生素组降低的程度最为明显，肉眼评分的顺序为对照组>抗生素组>PACT组>PACT+抗生素组(图2，D)。与治疗前相比，对照组的肉眼评分较前增加(p<0.05)，抗生素组的评分较前稍减低，PACT+抗生素组和PACT组的肉眼评分较前明显减低，其中以PACT+抗生素组减轻最为明显(p<0.01)。
X线检测：在治疗前、治疗2周后和治疗4周后分别用X线拍摄胫骨正侧位照片，观察胫骨的情况并进行放射学分析。用改良Norden骨髓炎评分法对骨髓炎感染的严重程度进行分级，0-7分代表骨髓炎从无至最严重程度。改良Norden骨髓炎评分包含4个指标分别为死骨形成、骨质破坏、骨质增生及软组织炎性包块影，每个指标都按照有、可疑及无三个等级进行评分，具体标准如表3。
表3Norden骨髓炎评分
X线能够有效的评估骨质的感染程度，对骨质的破坏及炎症反应所导致的骨质增生的评价有一定的价值。对照组在5周后能够看到明显的骨质破坏、吸收，胫骨骨干的增粗，周围软组织的肿胀。抗生素组有骨质的破坏、骨质的增生硬化及死骨的形成，周围软组织也有重度的肿胀。PACT组和PACT+抗生素组均没有明显的骨质破坏及增生硬化，周围软组织也未见明显的肿胀，其中，PACT+抗生素组骨缺损倾向于愈合(图2，C)。各组的Norden评分在治疗前未见明显的差异(p>0.05)。2周后及5周后，各治疗组的Norden评分与对照组相比均有减低，其中以PACT+抗生素组减轻最为明显，PACT组次之。与治疗前相比，对照组的Norden评分明显增加(p<0.05)，抗生素组的评分变化不明显，PACT+抗生素组和PACT组的Norden评分明显减低(p<0.001)，其中以PACT+抗生素组减轻最为明显(图2，E)。
细菌计数：各实验组分别在治疗前、治疗2周后和治疗4周后随机取出并处死实验动物，分离胫骨，以感染区为中心将胫骨切开，从胫骨起始骨隧道采集骨髓，加入无菌EP管中冷冻保存，并尽早进行骨髓组织研磨，用无菌生理盐水将研磨的原液稀释到10ml，采用稀释平板涂布法，取稀释液1ml于培养皿中，用涂布器涂匀，置于37℃孵育24h，进行细菌计数。
治疗2周后，对照组的菌落数较之前无明显变化，各治疗组的菌落数均较之前有明显的减少，其中以PACT+抗生素组菌落数下降最为明显，PACT组次之(图3，A)。5周后，对照组的菌落数与治疗前相比稍增多，抗生素组的菌落数与治疗2周相比稍增多，而PACT+抗生素组和PACT组的菌落数较治疗前明显减低(P<0.001)，其中以PACT+抗生素组菌落数减少的最多，PACT组次之(图3，B)。
组织学检查：各实验组分别在治疗前、治疗2周、5周后随机取出并处死实验动物，取出胫骨，将胫骨近端标本固定在中性福尔马林缓冲溶液中，用10％乙二胺四乙酸脱钙，嵌入石蜡中，切成6μm纵断面。切片用苏木精-伊红进行染色。Smeltzer评分系统包含4个指标，通过评估骨内急性炎症、骨内慢性炎症、骨膜炎症及骨坏死来进行组织病理学分级，每个参数分5级(0-4分)。各组织病理学参数的总和计为总组织学评分，其总分16分，≧8分定义为明显感染。评分是由一位独立的、研究外、双盲的研究人员对所有切片进行评分。具体评分标准如表4。
表4 Smeltzer组织学评分
组织学能够在细胞分子层面评价PACT对骨髓炎的治疗效果。实验结果显示对照组可见明显的骨内炎症，胫骨骨骺破坏并伴有严重脓肿。抗生素组在治疗2周后有中度的骨内和骨膜炎症，轻度的骨破坏。而PACT组和PACT+抗生素组在治疗2周后仅有轻微的骨内和骨膜炎症，坏死仅见于骨隧道周围，无骨破坏或关节破坏。5周后，对照组和抗生素组可见明显的骨内炎症及骨质的破坏，骨组织周围有大量的炎性细胞浸润，部分标本内可见成骨细胞无规则的排列(图4，A)。而PACT组和PACT+抗生素组炎性细胞较前明显减少，骨缺损由新生骨填充，可见规则的骨细胞，有时还可见新生的成骨细胞。通过对各组骨的标本进行评分，结果显示对照组的Smeltzer评分最高，PACT+抗生素组的评分最低。PACT+抗生素组和PACT组与对照组和抗生素组相比，均有明显的差异(p<0.01)(图4，B)。
降钙素原的检测：在造模前、治疗前以及治疗后的每周对各实验组兔子进行耳缘静脉采血(2ml)，离心(3000r/min)取血清，并将提取的血清放于1ml的无菌EP管中保存(-20℃)，ELISA检测试剂盒测定血清中降钙素原的含量。
降钙素原是检测体内炎症反应的有效指标。造模前各组降钙素原的含量都在正常范围，造模后1天及造模后1周各组的降钙素原含量都较之前有明显的升高(p<0.001)，但各组之间没有明显的差异(p>0.05)。随着治疗时间的延长，与对照组相比，各治疗组的降钙素原的含量均有明显的下降((p>0.05))，其中以PACT+抗生素组下降最为明显，PACT组次之(图5)。其中PACT+抗生素组和PACT组的含量随着治疗时间的增加在不断下降并维持在治疗前的正常水平。PACT+抗生素组在4周达到治疗前的正常水平，PACT组在5周达到正常水平。虽然抗生素组降钙素原的含量随着治疗时间的延长也有明显的下降，但一直没有下降到正常水平。
毒副作用检测：分别将处死的实验动物脏器(心、肝、脾、肺及肾)取出，去除血液后称重(精确到0.1g)，计算脏器指数(脏器质量/体重)。称重后分别取出相应组织，将其固定在中性福尔马林缓冲溶液中，固定48h后进行石蜡包埋、切片及染色。切片用苏木精-伊红染色。
PACT+抗生素组、PACT组及抗生素组，各脏器的脏器指数与对照组相比差异无统计学意义(p>0.05，表5)。各治疗组和对照组的脏器组织HE染色结果均没有见明显的病理损伤(图6)，说明新型光敏剂LD4对动物机体无明显的毒副作用。
表5各组实验动物的脏器指数
统计学分析：SPSS19.0软件进行数据处理。数据用平均数±标准差表示，多组间比较采用重复测量资料的方差分析。同一时刻不同组间比较采用单因素方差分析，多重比较用最小显著差异法(LSD)T检验。P<0.05认为有统计学意义。Origin7.0对数据进行图表制作。
Claims (2)
1.碱性氨基酸修饰氨基四苯基卟啉化合物在制备光动力治疗骨髓炎药物的应用，所述的碱性氨基酸修饰氨基四苯基卟啉化合物具有下述结构：
2.根据权利要求1所述的用途，其特征是所述的药物为体内光动力治疗药物。
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202210342791.0A CN114558136A (zh) | 2022-03-31 | 2022-03-31 | 碱性氨基酸修饰氨基四苯基卟啉化合物治疗骨髓炎的用途 |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202210342791.0A CN114558136A (zh) | 2022-03-31 | 2022-03-31 | 碱性氨基酸修饰氨基四苯基卟啉化合物治疗骨髓炎的用途 |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN114558136A true CN114558136A (zh) | 2022-05-31 |
Family
ID=81719366
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202210342791.0A Pending CN114558136A (zh) | 2022-03-31 | 2022-03-31 | 碱性氨基酸修饰氨基四苯基卟啉化合物治疗骨髓炎的用途 |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN114558136A (zh) |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103772399A (zh) * | 2014-02-21 | 2014-05-07 | 中国医学科学院生物医学工程研究所 | 碱性氨基酸修饰氨基四苯基卟啉化合物的制备方法及用途 |
| WO2016051361A1 (en) * | 2014-09-30 | 2016-04-07 | Biolitec Unternehmensbeteiligungs Ii Ag | Specifically meso-substituted porphyrins and chlorins for photodynamic therapy |
| CN107033153A (zh) * | 2017-06-05 | 2017-08-11 | 中国医学科学院生物医学工程研究所 | 鸟氨酸修饰氨基四苯基卟啉化合物及用途 |
-
2022
- 2022-03-31 CN CN202210342791.0A patent/CN114558136A/zh active Pending
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103772399A (zh) * | 2014-02-21 | 2014-05-07 | 中国医学科学院生物医学工程研究所 | 碱性氨基酸修饰氨基四苯基卟啉化合物的制备方法及用途 |
| WO2016051361A1 (en) * | 2014-09-30 | 2016-04-07 | Biolitec Unternehmensbeteiligungs Ii Ag | Specifically meso-substituted porphyrins and chlorins for photodynamic therapy |
| CN107033153A (zh) * | 2017-06-05 | 2017-08-11 | 中国医学科学院生物医学工程研究所 | 鸟氨酸修饰氨基四苯基卟啉化合物及用途 |
Non-Patent Citations (1)
| Title |
|---|
| SHUAI MENG ET AL: "Synthesis, characterization and in vitro photodynamic antimicrobial activity of basic amino acideporphyrin conjugates" * |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Lutwick et al. | Deep infections from Petriellidium boydii treated with miconazole | |
| JP6703040B2 (ja) | 角膜創傷治癒および眼表面疾患のためのヒスタチン | |
| Orenstein et al. | Effects of mast cell modulation on early host response to implanted synthetic meshes | |
| KR102597594B1 (ko) | 오가노이드의 생체 이식용 조성물 | |
| ES2841742T3 (es) | Nuevo uso de derivados triazolo(4,5-d)pirimidina para la prevención y tratamiento de infección bacteriana | |
| CN107157950A (zh) | 一种白蛋白纳米粒及其制备方法和用途 | |
| Hill et al. | Squamous cell carcinoma with hepatic metastasis in a saltwater crocodile (Crocodylus porosus). | |
| CN109529045B (zh) | 利福霉素-喹嗪酮偶联分子及其药学上可接受的盐的应用 | |
| CN113234125B (zh) | 自组装多肽、多肽水凝胶及其制备方法和用途 | |
| CN114558136A (zh) | 碱性氨基酸修饰氨基四苯基卟啉化合物治疗骨髓炎的用途 | |
| JP6863973B2 (ja) | タウロリジンの皮膚浸透製剤 | |
| EP1677811B1 (en) | Medicament on the basis of honey, preparation and use thereof | |
| CN111518187A (zh) | 抗菌肽dn6nh2及其应用 | |
| JP2021502410A (ja) | 細胞内細菌感染症治療用組成物 | |
| CN111920813A (zh) | 6-乙氧基血根碱在制备抗肿瘤药物中的应用 | |
| CN113398275A (zh) | 一种用于光动力治疗的细菌载体及其制备方法与应用 | |
| CN111838075A (zh) | 高毒肺炎克雷伯菌的液体气溶胶肺递送小鼠感染模型 | |
| CN116942696B (zh) | 亚铁离子在制备烫伤感染治疗药物和烫伤护理用品中的应用 | |
| WO2020069488A1 (en) | Compositions and methods for drug delivery | |
| US10398664B2 (en) | Methods of diagnosing and treating infected implants | |
| KR100674604B1 (ko) | 싸이클릭다이펩타이드를 주성분으로 하는 방사선 보호제 | |
| CN115227817A (zh) | 一种激光驱动的bbr@sp生物活性水凝胶、制备方法及用途 | |
| CN110559451B (zh) | 一种纳米药物及其制备方法和应用 | |
| CN111527085B (zh) | 用于细菌感染局部治疗的5-(2,5-双(4-氯-2-异丙基苯基)噻吩-3-基)-1h-四唑 | |
| Pang et al. | Use of a pH-responsive imatinib mesylate sustained-release hydrogel for the treatment of tendon adhesion by inhibiting PDGFRβ/CLDN1 pathway |
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 | ||
| WD01 | Invention patent application deemed withdrawn after publication |
Application publication date: 20220531 |
|
| WD01 | Invention patent application deemed withdrawn after publication |