WO1988000195A1 - Misakinolide compositions and their derivatives - Google Patents
Misakinolide compositions and their derivatives Download PDFInfo
- Publication number
- WO1988000195A1 WO1988000195A1 PCT/US1987/001565 US8701565W WO8800195A1 WO 1988000195 A1 WO1988000195 A1 WO 1988000195A1 US 8701565 W US8701565 W US 8701565W WO 8800195 A1 WO8800195 A1 WO 8800195A1
- Authority
- WO
- WIPO (PCT)
- Prior art keywords
- compositions
- contacting
- amount
- effective
- inhibiting
- Prior art date
Links
- 239000000203 mixture Substances 0.000 title claims abstract description 112
- KOMVIEGHPLWKNT-UHFFFAOYSA-N misakinolide-A Natural products C1C(OC)CC(C)OC1CCC(C)C(O)C(C)C1C(C)C(O)CC(O)C(C)C(OC)CC(CC=C2)OC2CC(O)CC=C(C)C(=O)O1 KOMVIEGHPLWKNT-UHFFFAOYSA-N 0.000 title abstract description 16
- 238000000034 method Methods 0.000 claims abstract description 48
- 206010028980 Neoplasm Diseases 0.000 claims abstract description 32
- 241000700605 Viruses Species 0.000 claims abstract description 28
- 230000000259 anti-tumor effect Effects 0.000 claims abstract description 27
- 230000002401 inhibitory effect Effects 0.000 claims abstract description 27
- 230000000840 anti-viral effect Effects 0.000 claims abstract description 26
- 241000233866 Fungi Species 0.000 claims abstract description 24
- 241000243142 Porifera Species 0.000 claims abstract description 15
- 230000008569 process Effects 0.000 claims abstract description 13
- 241001521381 Theonella Species 0.000 claims abstract description 7
- 125000004423 acyloxy group Chemical group 0.000 claims abstract description 6
- 125000003545 alkoxy group Chemical group 0.000 claims abstract description 6
- 239000012871 anti-fungal composition Substances 0.000 claims abstract description 6
- 229910052739 hydrogen Inorganic materials 0.000 claims abstract description 5
- 239000001257 hydrogen Substances 0.000 claims abstract description 5
- 125000004435 hydrogen atom Chemical class [H]* 0.000 claims abstract 2
- 125000002887 hydroxy group Chemical group [H]O* 0.000 claims abstract 2
- 230000000843 anti-fungal effect Effects 0.000 claims description 14
- 230000012010 growth Effects 0.000 claims description 14
- 239000000284 extract Substances 0.000 claims description 13
- 229940121375 antifungal agent Drugs 0.000 claims description 12
- 239000002904 solvent Substances 0.000 claims description 12
- 239000003085 diluting agent Substances 0.000 claims description 10
- 239000003937 drug carrier Substances 0.000 claims description 10
- 231100000252 nontoxic Toxicity 0.000 claims description 10
- 230000003000 nontoxic effect Effects 0.000 claims description 10
- 230000002829 reductive effect Effects 0.000 claims description 9
- 239000004480 active ingredient Substances 0.000 claims description 7
- 239000000243 solution Substances 0.000 claims description 7
- 206010006895 Cachexia Diseases 0.000 claims description 5
- 239000002253 acid Substances 0.000 claims description 4
- 239000012670 alkaline solution Substances 0.000 claims description 4
- 239000003960 organic solvent Substances 0.000 claims description 4
- 238000002560 therapeutic procedure Methods 0.000 claims description 3
- 239000003120 macrolide antibiotic agent Substances 0.000 abstract description 31
- 210000004027 cell Anatomy 0.000 description 41
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 21
- XEKOWRVHYACXOJ-UHFFFAOYSA-N Ethyl acetate Chemical compound CCOC(C)=O XEKOWRVHYACXOJ-UHFFFAOYSA-N 0.000 description 21
- CSCPPACGZOOCGX-UHFFFAOYSA-N Acetone Chemical compound CC(C)=O CSCPPACGZOOCGX-UHFFFAOYSA-N 0.000 description 18
- 241000282414 Homo sapiens Species 0.000 description 18
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 16
- PEDCQBHIVMGVHV-UHFFFAOYSA-N Glycerine Chemical compound OCC(O)CO PEDCQBHIVMGVHV-UHFFFAOYSA-N 0.000 description 15
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 description 15
- 238000003556 assay Methods 0.000 description 15
- 201000010099 disease Diseases 0.000 description 15
- 241000124008 Mammalia Species 0.000 description 12
- 241000196324 Embryophyta Species 0.000 description 11
- RTZKZFJDLAIYFH-UHFFFAOYSA-N Diethyl ether Chemical compound CCOCC RTZKZFJDLAIYFH-UHFFFAOYSA-N 0.000 description 10
- 239000002609 medium Substances 0.000 description 9
- 230000003612 virological effect Effects 0.000 description 9
- 150000001875 compounds Chemical class 0.000 description 8
- 230000000694 effects Effects 0.000 description 8
- 239000000417 fungicide Substances 0.000 description 8
- 229940079593 drug Drugs 0.000 description 7
- 239000003814 drug Substances 0.000 description 7
- 208000015181 infectious disease Diseases 0.000 description 7
- 241000222122 Candida albicans Species 0.000 description 6
- IAZDPXIOMUYVGZ-UHFFFAOYSA-N Dimethylsulphoxide Chemical compound CS(C)=O IAZDPXIOMUYVGZ-UHFFFAOYSA-N 0.000 description 6
- 208000031888 Mycoses Diseases 0.000 description 6
- LEHBURLTIWGHEM-UHFFFAOYSA-N pyridinium chlorochromate Chemical compound [O-][Cr](Cl)(=O)=O.C1=CC=[NH+]C=C1 LEHBURLTIWGHEM-UHFFFAOYSA-N 0.000 description 6
- 108091003079 Bovine Serum Albumin Proteins 0.000 description 5
- 241000282412 Homo Species 0.000 description 5
- 239000003810 Jones reagent Substances 0.000 description 5
- 241000711975 Vesicular stomatitis virus Species 0.000 description 5
- 239000012091 fetal bovine serum Substances 0.000 description 5
- 238000012423 maintenance Methods 0.000 description 5
- 238000002360 preparation method Methods 0.000 description 5
- 239000000047 product Substances 0.000 description 5
- 210000004881 tumor cell Anatomy 0.000 description 5
- 206010006187 Breast cancer Diseases 0.000 description 4
- QWOJMRHUQHTCJG-UHFFFAOYSA-N CC([CH2-])=O Chemical compound CC([CH2-])=O QWOJMRHUQHTCJG-UHFFFAOYSA-N 0.000 description 4
- HEDRZPFGACZZDS-UHFFFAOYSA-N Chloroform Chemical compound ClC(Cl)Cl HEDRZPFGACZZDS-UHFFFAOYSA-N 0.000 description 4
- 241000700588 Human alphaherpesvirus 1 Species 0.000 description 4
- VEXZGXHMUGYJMC-UHFFFAOYSA-N Hydrochloric acid Chemical compound Cl VEXZGXHMUGYJMC-UHFFFAOYSA-N 0.000 description 4
- 241001465754 Metazoa Species 0.000 description 4
- 241000985542 Penicillium atrovenetum Species 0.000 description 4
- UHOVQNZJYSORNB-MZWXYZOWSA-N benzene-d6 Chemical compound [2H]C1=C([2H])C([2H])=C([2H])C([2H])=C1[2H] UHOVQNZJYSORNB-MZWXYZOWSA-N 0.000 description 4
- 201000011510 cancer Diseases 0.000 description 4
- 239000003153 chemical reaction reagent Substances 0.000 description 4
- 238000011534 incubation Methods 0.000 description 4
- -1 mycotic disease Chemical compound 0.000 description 4
- 230000009467 reduction Effects 0.000 description 4
- 238000006722 reduction reaction Methods 0.000 description 4
- 239000000126 substance Substances 0.000 description 4
- 238000012360 testing method Methods 0.000 description 4
- ZWEHNKRNPOVVGH-UHFFFAOYSA-N 2-Butanone Chemical compound CCC(C)=O ZWEHNKRNPOVVGH-UHFFFAOYSA-N 0.000 description 3
- YMWUJEATGCHHMB-UHFFFAOYSA-N Dichloromethane Chemical compound ClCCl YMWUJEATGCHHMB-UHFFFAOYSA-N 0.000 description 3
- 239000006145 Eagle's minimal essential medium Substances 0.000 description 3
- 206010017533 Fungal infection Diseases 0.000 description 3
- 241000238631 Hexapoda Species 0.000 description 3
- 206010058467 Lung neoplasm malignant Diseases 0.000 description 3
- KWYUFKZDYYNOTN-UHFFFAOYSA-M Potassium hydroxide Chemical compound [OH-].[K+] KWYUFKZDYYNOTN-UHFFFAOYSA-M 0.000 description 3
- YXFVVABEGXRONW-UHFFFAOYSA-N Toluene Chemical compound CC1=CC=CC=C1 YXFVVABEGXRONW-UHFFFAOYSA-N 0.000 description 3
- 208000029742 colonic neoplasm Diseases 0.000 description 3
- 231100000135 cytotoxicity Toxicity 0.000 description 3
- 230000003013 cytotoxicity Effects 0.000 description 3
- 230000000855 fungicidal effect Effects 0.000 description 3
- 238000000338 in vitro Methods 0.000 description 3
- 230000005764 inhibitory process Effects 0.000 description 3
- 208000032839 leukemia Diseases 0.000 description 3
- 239000012280 lithium aluminium hydride Substances 0.000 description 3
- 201000005296 lung carcinoma Diseases 0.000 description 3
- 229920000609 methyl cellulose Polymers 0.000 description 3
- 239000001923 methylcellulose Substances 0.000 description 3
- 244000005700 microbiome Species 0.000 description 3
- 230000004048 modification Effects 0.000 description 3
- 238000012986 modification Methods 0.000 description 3
- 239000003921 oil Substances 0.000 description 3
- 230000036961 partial effect Effects 0.000 description 3
- 230000002265 prevention Effects 0.000 description 3
- 239000002689 soil Substances 0.000 description 3
- 210000001519 tissue Anatomy 0.000 description 3
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 3
- 229920001817 Agar Polymers 0.000 description 2
- 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 2
- 229930182566 Gentamicin Natural products 0.000 description 2
- WQZGKKKJIJFFOK-GASJEMHNSA-N Glucose Natural products OC[C@H]1OC(O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-GASJEMHNSA-N 0.000 description 2
- 229910010084 LiAlH4 Inorganic materials 0.000 description 2
- KDLHZDBZIXYQEI-UHFFFAOYSA-N Palladium Chemical compound [Pd] KDLHZDBZIXYQEI-UHFFFAOYSA-N 0.000 description 2
- BELBBZDIHDAJOR-UHFFFAOYSA-N Phenolsulfonephthalein Chemical compound C1=CC(O)=CC=C1C1(C=2C=CC(O)=CC=2)C2=CC=CC=C2S(=O)(=O)O1 BELBBZDIHDAJOR-UHFFFAOYSA-N 0.000 description 2
- JUJWROOIHBZHMG-UHFFFAOYSA-N Pyridine Chemical compound C1=CC=NC=C1 JUJWROOIHBZHMG-UHFFFAOYSA-N 0.000 description 2
- QAOWNCQODCNURD-UHFFFAOYSA-N Sulfuric acid Chemical compound OS(O)(=O)=O QAOWNCQODCNURD-UHFFFAOYSA-N 0.000 description 2
- JXLYSJRDGCGARV-WWYNWVTFSA-N Vinblastine Natural products O=C(O[C@H]1[C@](O)(C(=O)OC)[C@@H]2N(C)c3c(cc(c(OC)c3)[C@]3(C(=O)OC)c4[nH]c5c(c4CCN4C[C@](O)(CC)C[C@H](C3)C4)cccc5)[C@@]32[C@H]2[C@@]1(CC)C=CCN2CC3)C JXLYSJRDGCGARV-WWYNWVTFSA-N 0.000 description 2
- 239000008272 agar Substances 0.000 description 2
- 239000003429 antifungal agent Substances 0.000 description 2
- 239000002246 antineoplastic agent Substances 0.000 description 2
- 239000003443 antiviral agent Substances 0.000 description 2
- 201000008274 breast adenocarcinoma Diseases 0.000 description 2
- 229940095731 candida albicans Drugs 0.000 description 2
- 125000004432 carbon atom Chemical group C* 0.000 description 2
- 238000010531 catalytic reduction reaction Methods 0.000 description 2
- 238000004113 cell culture Methods 0.000 description 2
- 238000006243 chemical reaction Methods 0.000 description 2
- 238000001816 cooling Methods 0.000 description 2
- 230000006378 damage Effects 0.000 description 2
- 239000008121 dextrose Substances 0.000 description 2
- 238000011156 evaluation Methods 0.000 description 2
- 238000005194 fractionation Methods 0.000 description 2
- 239000000499 gel Substances 0.000 description 2
- 239000001963 growth medium Substances 0.000 description 2
- 150000002431 hydrogen Chemical class 0.000 description 2
- 125000004356 hydroxy functional group Chemical group O* 0.000 description 2
- 239000002054 inoculum Substances 0.000 description 2
- 238000002955 isolation Methods 0.000 description 2
- 230000002147 killing effect Effects 0.000 description 2
- 210000004072 lung Anatomy 0.000 description 2
- 238000010899 nucleation Methods 0.000 description 2
- 229960003531 phenolsulfonphthalein Drugs 0.000 description 2
- 230000002685 pulmonary effect Effects 0.000 description 2
- 238000011160 research Methods 0.000 description 2
- 239000012449 sabouraud dextrose agar Substances 0.000 description 2
- 210000002966 serum Anatomy 0.000 description 2
- 239000007858 starting material Substances 0.000 description 2
- RJVBVECTCMRNFG-ANKJNSLFSA-N swinholide a Chemical compound C1[C@H](OC)C[C@H](C)O[C@H]1CC[C@H](C)[C@H](O)[C@H](C)[C@@H]1[C@@H](C)[C@H](O)C[C@H](O)[C@H](C)[C@@H](OC)C[C@H](CC=C2)O[C@@H]2C[C@@H](O)C/C=C(\C)/C=C/C(=O)O[C@H]([C@@H](C)[C@@H](O)[C@@H](C)CC[C@@H]2O[C@@H](C)C[C@H](C2)OC)[C@@H](C)[C@H](O)C[C@H](O)[C@H](C)[C@@H](OC)C[C@H](CC=C2)O[C@@H]2C[C@@H](O)C/C=C(\C)/C=C/C(=O)O1 RJVBVECTCMRNFG-ANKJNSLFSA-N 0.000 description 2
- GDACDJNQZCXLNU-UHFFFAOYSA-N swinholide-A Natural products C1C(OC)CC(C)OC1CCC(C)C(O)C(C)C1C(C)C(O)CC(O)C(C)C(OC)CC(CC=C2)OC2CC(O)CC=C(C)C=CC(=O)O1 GDACDJNQZCXLNU-UHFFFAOYSA-N 0.000 description 2
- 229960003048 vinblastine Drugs 0.000 description 2
- 229960004528 vincristine Drugs 0.000 description 2
- OGWKCGZFUXNPDA-XQKSVPLYSA-N vincristine Chemical compound C([N@]1C[C@@H](C[C@]2(C(=O)OC)C=3C(=CC4=C([C@]56[C@H]([C@@]([C@H](OC(C)=O)[C@]7(CC)C=CCN([C@H]67)CC5)(O)C(=O)OC)N4C=O)C=3)OC)C[C@@](C1)(O)CC)CC1=C2NC2=CC=CC=C12 OGWKCGZFUXNPDA-XQKSVPLYSA-N 0.000 description 2
- OGWKCGZFUXNPDA-UHFFFAOYSA-N vincristine Natural products C1C(CC)(O)CC(CC2(C(=O)OC)C=3C(=CC4=C(C56C(C(C(OC(C)=O)C7(CC)C=CCN(C67)CC5)(O)C(=O)OC)N4C=O)C=3)OC)CN1CCC1=C2NC2=CC=CC=C12 OGWKCGZFUXNPDA-UHFFFAOYSA-N 0.000 description 2
- BLSQLHNBWJLIBQ-OZXSUGGESA-N (2R,4S)-terconazole Chemical compound C1CN(C(C)C)CCN1C(C=C1)=CC=C1OC[C@@H]1O[C@@](CN2N=CN=C2)(C=2C(=CC(Cl)=CC=2)Cl)OC1 BLSQLHNBWJLIBQ-OZXSUGGESA-N 0.000 description 1
- 238000001644 13C nuclear magnetic resonance spectroscopy Methods 0.000 description 1
- 238000005160 1H NMR spectroscopy Methods 0.000 description 1
- PKDBCJSWQUOKDO-UHFFFAOYSA-M 2,3,5-triphenyltetrazolium chloride Chemical compound [Cl-].C1=CC=CC=C1C(N=[N+]1C=2C=CC=CC=2)=NN1C1=CC=CC=C1 PKDBCJSWQUOKDO-UHFFFAOYSA-M 0.000 description 1
- RBTBFTRPCNLSDE-UHFFFAOYSA-N 3,7-bis(dimethylamino)phenothiazin-5-ium Chemical compound C1=CC(N(C)C)=CC2=[S+]C3=CC(N(C)C)=CC=C3N=C21 RBTBFTRPCNLSDE-UHFFFAOYSA-N 0.000 description 1
- DENKGPBHLYFNGK-UHFFFAOYSA-N 4-bromobenzoyl chloride Chemical compound ClC(=O)C1=CC=C(Br)C=C1 DENKGPBHLYFNGK-UHFFFAOYSA-N 0.000 description 1
- 206010005098 Blastomycosis Diseases 0.000 description 1
- 208000026310 Breast neoplasm Diseases 0.000 description 1
- 206010007134 Candida infections Diseases 0.000 description 1
- 241000282552 Chlorocebus aethiops Species 0.000 description 1
- 201000007336 Cryptococcosis Diseases 0.000 description 1
- 241000221204 Cryptococcus neoformans Species 0.000 description 1
- 241000192700 Cyanobacteria Species 0.000 description 1
- 239000012591 Dulbecco’s Phosphate Buffered Saline Substances 0.000 description 1
- 208000009889 Herpes Simplex Diseases 0.000 description 1
- UFHFLCQGNIYNRP-UHFFFAOYSA-N Hydrogen Chemical compound [H][H] UFHFLCQGNIYNRP-UHFFFAOYSA-N 0.000 description 1
- XQFRJNBWHJMXHO-RRKCRQDMSA-N IDUR Chemical compound C1[C@H](O)[C@@H](CO)O[C@H]1N1C(=O)NC(=O)C(I)=C1 XQFRJNBWHJMXHO-RRKCRQDMSA-N 0.000 description 1
- NTIZESTWPVYFNL-UHFFFAOYSA-N Methyl isobutyl ketone Chemical compound CC(C)CC(C)=O NTIZESTWPVYFNL-UHFFFAOYSA-N 0.000 description 1
- UIHCLUNTQKBZGK-UHFFFAOYSA-N Methyl isobutyl ketone Natural products CCC(C)C(C)=O UIHCLUNTQKBZGK-UHFFFAOYSA-N 0.000 description 1
- 241000208125 Nicotiana Species 0.000 description 1
- 235000002637 Nicotiana tabacum Nutrition 0.000 description 1
- 206010035148 Plague Diseases 0.000 description 1
- 239000004793 Polystyrene Substances 0.000 description 1
- 108010020147 Protein Corona Proteins 0.000 description 1
- NPXOKRUENSOPAO-UHFFFAOYSA-N Raney nickel Chemical compound [Al].[Ni] NPXOKRUENSOPAO-UHFFFAOYSA-N 0.000 description 1
- 240000004808 Saccharomyces cerevisiae Species 0.000 description 1
- 235000014680 Saccharomyces cerevisiae Nutrition 0.000 description 1
- CYZZFSCFKCNCLO-UHFFFAOYSA-N Scytophycin A Natural products COC1CC(OC)C(C)C(C(C)C(O)C(C)CCC(O)C(C)C(C(C)C=CN(C)C=O)OC)OC(=O)C=CC(C)=CCC(O)CC(C=CC2)OC2CC(OC)C21CO2 CYZZFSCFKCNCLO-UHFFFAOYSA-N 0.000 description 1
- LGYQYCWMQRMLJJ-DAAXTDNESA-N Scytophycin B Chemical compound C([C@@]12[C@@H](OC)C[C@H]3O[C@@H](C=CC3)C[C@@H](O)C/C=C(\C)/C=C/C(=O)O[C@@H]([C@H]([C@H](OC)C[C@@H]2OC)C)[C@@H](C)[C@@H](O)[C@@H](C)CCC(=O)[C@H](C)[C@@H]([C@H](C)\C=C\N(C)C=O)OC)O1 LGYQYCWMQRMLJJ-DAAXTDNESA-N 0.000 description 1
- LGYQYCWMQRMLJJ-UHFFFAOYSA-N Scytophycin B Natural products COC1CC(OC)C(C)C(C(C)C(O)C(C)CCC(=O)C(C)C(C(C)C=CN(C)C=O)OC)OC(=O)C=CC(C)=CCC(O)CC(C=CC2)OC2CC(OC)C21CO2 LGYQYCWMQRMLJJ-UHFFFAOYSA-N 0.000 description 1
- VYPSYNLAJGMNEJ-UHFFFAOYSA-N Silicium dioxide Chemical compound O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 description 1
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 1
- 241001521370 Theonella swinhoei Species 0.000 description 1
- 208000002474 Tinea Diseases 0.000 description 1
- 241000893966 Trichophyton verrucosum Species 0.000 description 1
- 229920004890 Triton X-100 Polymers 0.000 description 1
- 239000013504 Triton X-100 Substances 0.000 description 1
- 102000004142 Trypsin Human genes 0.000 description 1
- 108090000631 Trypsin Proteins 0.000 description 1
- 241000607479 Yersinia pestis Species 0.000 description 1
- 230000002159 abnormal effect Effects 0.000 description 1
- 239000003929 acidic solution Substances 0.000 description 1
- 208000009956 adenocarcinoma Diseases 0.000 description 1
- 125000000217 alkyl group Chemical group 0.000 description 1
- 230000000202 analgesic effect Effects 0.000 description 1
- 230000001032 anti-candidal effect Effects 0.000 description 1
- 239000007864 aqueous solution Substances 0.000 description 1
- 239000007900 aqueous suspension Substances 0.000 description 1
- 239000012298 atmosphere Substances 0.000 description 1
- 230000033228 biological regulation Effects 0.000 description 1
- 244000309466 calf Species 0.000 description 1
- 201000003984 candidiasis Diseases 0.000 description 1
- 125000002915 carbonyl group Chemical group [*:2]C([*:1])=O 0.000 description 1
- 150000001732 carboxylic acid derivatives Chemical class 0.000 description 1
- 239000000969 carrier Substances 0.000 description 1
- 230000015556 catabolic process Effects 0.000 description 1
- 239000003054 catalyst Substances 0.000 description 1
- 238000006555 catalytic reaction Methods 0.000 description 1
- 230000010261 cell growth Effects 0.000 description 1
- 210000003169 central nervous system Anatomy 0.000 description 1
- 239000013043 chemical agent Substances 0.000 description 1
- 239000007795 chemical reaction product Substances 0.000 description 1
- 238000002512 chemotherapy Methods 0.000 description 1
- 238000004587 chromatography analysis Methods 0.000 description 1
- 210000001072 colon Anatomy 0.000 description 1
- 201000010897 colon adenocarcinoma Diseases 0.000 description 1
- 239000012230 colorless oil Substances 0.000 description 1
- 239000013078 crystal Substances 0.000 description 1
- 238000006731 degradation reaction Methods 0.000 description 1
- 230000018044 dehydration Effects 0.000 description 1
- 238000006297 dehydration reaction Methods 0.000 description 1
- WGLUMOCWFMKWIL-UHFFFAOYSA-N dichloromethane;methanol Chemical compound OC.ClCCl WGLUMOCWFMKWIL-UHFFFAOYSA-N 0.000 description 1
- 238000009792 diffusion process Methods 0.000 description 1
- 239000012895 dilution Substances 0.000 description 1
- 238000010790 dilution Methods 0.000 description 1
- 208000035475 disorder Diseases 0.000 description 1
- 239000012153 distilled water Substances 0.000 description 1
- 241001493065 dsRNA viruses Species 0.000 description 1
- 235000013399 edible fruits Nutrition 0.000 description 1
- 238000000605 extraction Methods 0.000 description 1
- 235000013305 food Nutrition 0.000 description 1
- 238000007710 freezing Methods 0.000 description 1
- 230000008014 freezing Effects 0.000 description 1
- 230000002538 fungal effect Effects 0.000 description 1
- 229940089256 fungistat Drugs 0.000 description 1
- 210000001035 gastrointestinal tract Anatomy 0.000 description 1
- ZDXPYRJPNDTMRX-UHFFFAOYSA-N glutamine Natural products OC(=O)C(N)CCC(N)=O ZDXPYRJPNDTMRX-UHFFFAOYSA-N 0.000 description 1
- 230000009036 growth inhibition Effects 0.000 description 1
- 229910052736 halogen Inorganic materials 0.000 description 1
- 150000002367 halogens Chemical group 0.000 description 1
- 230000036541 health Effects 0.000 description 1
- 208000019622 heart disease Diseases 0.000 description 1
- 238000005984 hydrogenation reaction Methods 0.000 description 1
- 230000005918 in vitro anti-tumor Effects 0.000 description 1
- 150000002576 ketones Chemical class 0.000 description 1
- 230000000670 limiting effect Effects 0.000 description 1
- 239000007788 liquid Substances 0.000 description 1
- 208000037841 lung tumor Diseases 0.000 description 1
- GBMDVOWEEQVZKZ-UHFFFAOYSA-N methanol;hydrate Chemical compound O.OC GBMDVOWEEQVZKZ-UHFFFAOYSA-N 0.000 description 1
- 229960000907 methylthioninium chloride Drugs 0.000 description 1
- 210000004400 mucous membrane Anatomy 0.000 description 1
- CYZZFSCFKCNCLO-XMPFBODESA-N n-[(e,3r,4r,5s,9s,10s,11s)-6,10-dihydroxy-11-[(1s,3s,4s,5s,7r,8s,9r,12e,14e,17s,19r)-17-hydroxy-3,5,7-trimethoxy-8,14-dimethyl-11-oxospiro[10,23-dioxabicyclo[17.3.1]tricosa-12,14,20-triene-4,2'-oxirane]-9-yl]-4-methoxy-3,5,9-trimethyldodec-1-enyl]-n-methy Chemical compound C([C@@]12[C@@H](OC)C[C@H]3O[C@@H](C=CC3)C[C@@H](O)C/C=C(\C)/C=C/C(=O)O[C@@H]([C@H]([C@H](OC)C[C@@H]2OC)C)[C@@H](C)[C@@H](O)[C@@H](C)CCC(O)[C@H](C)[C@@H]([C@H](C)\C=C\N(C)C=O)OC)O1 CYZZFSCFKCNCLO-XMPFBODESA-N 0.000 description 1
- 201000009240 nasopharyngitis Diseases 0.000 description 1
- PGSADBUBUOPOJS-UHFFFAOYSA-N neutral red Chemical compound Cl.C1=C(C)C(N)=CC2=NC3=CC(N(C)C)=CC=C3N=C21 PGSADBUBUOPOJS-UHFFFAOYSA-N 0.000 description 1
- 150000002894 organic compounds Chemical class 0.000 description 1
- 239000012044 organic layer Substances 0.000 description 1
- 239000012074 organic phase Substances 0.000 description 1
- 239000007800 oxidant agent Substances 0.000 description 1
- 230000003647 oxidation Effects 0.000 description 1
- 238000007254 oxidation reaction Methods 0.000 description 1
- 230000001590 oxidative effect Effects 0.000 description 1
- 239000012466 permeate Substances 0.000 description 1
- 239000012071 phase Substances 0.000 description 1
- 230000007505 plaque formation Effects 0.000 description 1
- 239000004033 plastic Substances 0.000 description 1
- 229920003023 plastic Polymers 0.000 description 1
- BASFCYQUMIYNBI-UHFFFAOYSA-N platinum Chemical compound [Pt] BASFCYQUMIYNBI-UHFFFAOYSA-N 0.000 description 1
- 229920002223 polystyrene Polymers 0.000 description 1
- 239000013641 positive control Substances 0.000 description 1
- 230000003449 preventive effect Effects 0.000 description 1
- 150000003138 primary alcohols Chemical group 0.000 description 1
- UMJSCPRVCHMLSP-UHFFFAOYSA-N pyridine Natural products COC1=CC=CN=C1 UMJSCPRVCHMLSP-UHFFFAOYSA-N 0.000 description 1
- 239000002994 raw material Substances 0.000 description 1
- 239000001044 red dye Substances 0.000 description 1
- 230000002441 reversible effect Effects 0.000 description 1
- 239000000741 silica gel Substances 0.000 description 1
- 229910002027 silica gel Inorganic materials 0.000 description 1
- 210000003491 skin Anatomy 0.000 description 1
- 239000011780 sodium chloride Substances 0.000 description 1
- 238000000638 solvent extraction Methods 0.000 description 1
- 230000003595 spectral effect Effects 0.000 description 1
- 230000007480 spreading Effects 0.000 description 1
- 208000003265 stomatitis Diseases 0.000 description 1
- 239000006228 supernatant Substances 0.000 description 1
- 208000024891 symptom Diseases 0.000 description 1
- 230000001225 therapeutic effect Effects 0.000 description 1
- 230000008467 tissue growth Effects 0.000 description 1
- JOXIMZWYDAKGHI-UHFFFAOYSA-N toluene-4-sulfonic acid Chemical compound CC1=CC=C(S(O)(=O)=O)C=C1 JOXIMZWYDAKGHI-UHFFFAOYSA-N 0.000 description 1
- 231100000331 toxic Toxicity 0.000 description 1
- 230000002588 toxic effect Effects 0.000 description 1
- 239000003053 toxin Substances 0.000 description 1
- 231100000765 toxin Toxicity 0.000 description 1
- 108700012359 toxins Proteins 0.000 description 1
- 238000012546 transfer Methods 0.000 description 1
- 239000012588 trypsin Substances 0.000 description 1
- 239000006150 trypticase soy agar Substances 0.000 description 1
- 230000004614 tumor growth Effects 0.000 description 1
- 241000712461 unidentified influenza virus Species 0.000 description 1
- 208000019553 vascular disease Diseases 0.000 description 1
- 235000013311 vegetables Nutrition 0.000 description 1
- 208000005925 vesicular stomatitis Diseases 0.000 description 1
- JXLYSJRDGCGARV-XQKSVPLYSA-N vincaleukoblastine Chemical compound C([C@@H](C[C@]1(C(=O)OC)C=2C(=CC3=C([C@]45[C@H]([C@@]([C@H](OC(C)=O)[C@]6(CC)C=CCN([C@H]56)CC4)(O)C(=O)OC)N3C)C=2)OC)C[C@@](C2)(O)CC)N2CCC2=C1NC1=CC=CC=C21 JXLYSJRDGCGARV-XQKSVPLYSA-N 0.000 description 1
- 230000000007 visual effect Effects 0.000 description 1
- 238000005406 washing Methods 0.000 description 1
- 208000016261 weight loss Diseases 0.000 description 1
- 230000004580 weight loss Effects 0.000 description 1
Classifications
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07D—HETEROCYCLIC COMPOUNDS
- C07D493/00—Heterocyclic compounds containing oxygen atoms as the only ring hetero atoms in the condensed system
- C07D493/12—Heterocyclic compounds containing oxygen atoms as the only ring hetero atoms in the condensed system in which the condensed system contains three hetero rings
- C07D493/18—Bridged systems
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P35/00—Antineoplastic agents
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07D—HETEROCYCLIC COMPOUNDS
- C07D493/00—Heterocyclic compounds containing oxygen atoms as the only ring hetero atoms in the condensed system
- C07D493/22—Heterocyclic compounds containing oxygen atoms as the only ring hetero atoms in the condensed system in which the condensed system contains four or more hetero rings
Definitions
- This application is a continuation-in-part of Serial No. 879,134 filed June 26, 1986.
- This invention relates to new organic compositions which have useful antitumor, antiviral and antifungal activity. Additionally and particularly, this invention relates to new antitumor, antiviral and antifungal misakinolide and derivative compositions derived from marine organisms, i.e., sponges of the genus Theonella and their methods of use.
- tumors are common in a variety of mammals and the prevention, control of the growth and regression of tumors in mammals is important to man.
- the term tumor refers to abnormal masses of new tissue growth which is discordant with the economy of the tissue of origin or of the host's body as a whole.
- Cancerous cachexia refers to the symptomatic discomfort that accompanies the infliction of a mammal with a tumor. These symptoms include weakened condition of the inflicted mammal as evidenced by, for example, weight loss. The seriousness of cancer is well know, e.g., cancer is second only to heart and vascular diseases as a cause of death in man.
- Viral diseases contribute to inflictions in humans including common colds, herpes and cancer and the importance of their control is obvious. Also important is control of viral diseases in animals for economic reasons as well as the ability of such animals to become virus reservoirs or carriers which facilitate the spreading of viral diseases to humans. Viral plant diseases have been known to have a disruptive effect on the cultivation of fruit trees, tobacco, and various vegetables. Insect viral diseases are also of interest because of the insects' ability to transfer viral diseases to humans.
- fungus may cause various diseases and infections in man including mycotic disease, e.g., pulmonary candidiasis and pulmonary blastomycosis.
- mycotic disease e.g., pulmonary candidiasis and pulmonary blastomycosis.
- Certain yeastlike organisms e.g., Cryptococcus neoformans, may cause serious infections of the central nervous system.
- More commonly known fungal infections in humans and mammals include ringworm, which are fungus infections of hair and nail areas, as well as resistant infections of the skin.
- Many other fungal infections inflict humans and mammals in the areas of skin, mucous membranes, intestinal tract, vaginal area and lungs.
- Plants are also attacked by various fungi. Damage caused by fungus infection to agriculture amounts to billions of dollars annually.
- Various inorganic and organic fungistats and fungicides have been tried but with limited success. It is of course important for the fungistat or fungicide to kill the fungi but not the plant and to leave no toxic residue on the food of the plant.
- Various methods have been utilized to combat fungus infection in agriculture including foliage fungicide by which method plants are coated with a preventive weather-resistant fungicide.
- Seed treatment and soil treatment are methods which require fungicides which are safe for seeds and resist degradation by soil and soil microorganisms.
- Chemotherapeutants are fungicides which permeate the plant to protect new growth or eliminate infections which have already occurred within the plant.
- Agricultural fungistats and fungicides and their application must also meet very stringent requirements and regulations, which have been promulgated, for example, in the United States.
- a potential source for antiviral compositions is marine plant and animal life and of particular interest are marine sponges. It has now been found that certain organic compounds derived from extracts of sponges of the genus Theonella, possess useful antitumor, antiviral and antifungal activity.
- R 1 , R 2 , R 3 R 4 R 5 and R 6 are the same or different and are a hydrogen, hydroxy, lower alkoxy, or lower acyloxy group.
- the invention also comprises reduced derivatives of these compositions wherein at least one of the double bonds of the composition described above according to formula I is reduced.
- the invention also comprises compositions formed by the treatment of any of the compositions of formula I and their reduced derivatives with oxidizing reagents, such as Jones reagent or pyridinium chlorochromate (PCC); with alkaline solutions; and with acidic solution.
- oxidizing reagents such as Jones reagent or pyridinium chlorochromate (PCC)
- PCC pyridinium chlorochromate
- compositions according to formula I as well as, their reduced derivatives and derivatives formed by treatment with Jones reagent , alkal ine or acid soluti ons will col lectively be referred to, hereinafter, as "macrolide compositions of the invention.”
- the composition is substantially pure.
- the lower alkoxy and acyloxy groups have from 1 to 5 carbon atoms.
- the invention comprises dimeric compositions of formulae II and ill:
- the invention also comprises an antitumor, antiviral or antifungal composition comprising, as active ingredient, an effective antitumor, antiviral or antifungal amount, respectively, of one or more macrolide compositions of the invention and a non-toxic pharmaceutically acceptable carrier or diluent.
- the invention also comprises a process to produce the macrolide compositions of the invention.
- the process comprises the steps of collecting marine sponge of the genus Theonella, contacting the marine sponge with a suitable organic solvent system to obtain an extract; fractionating the extract; and isolating a macrolide composition of the invention from the fractionated extract.
- the invention further comprises a method for inhibiting tumors comprising contacting tumor cells with an effective antitumor amount of one or more macrolide compositions of the invention.
- the invention further comprises method for inhibiting viruses comprising contacting viruses with an effective antiviral amount of one or more macrolide compositions of the invention.
- the invention further comprises a method for inhibiting the growth of or killing fungi comprising contacting fungi with an effective antifungal amount of one or more macrolide compositions of the invention.
- the invention comprises macrolide compositions of the general form
- R 1 R 2 , R 3 R 4 R 5 and R 6 are the same or different and are a hydrogen, hydroxy, lower alkoxy, or lower acyloxy group.
- the invention also comprises reduced derivatives of these compositions wherein at least one of the doubl'e bonds of the composition described above according to formula I is reduced.
- the composition is substantially pure.
- the lower alkoxy or acyloxy groups have from 1 to 5 carbon atoms.
- the invention comprises compositions of the formulae II and III:
- an antitumor composition comprising as active ingredient an effective antitumor amount of one or more of the macrolide compositions of the invention and a non-toxic pharmaceutically acceptable carrier or diluent. While effective amount?, may vary, as conditions in which the antitumor compositions are used vary, a minimal dosage required for activity is generally between 0.01 and 10
- non-toxic pharmaceutically acceptable carriers or diluents include, but are not limited to, the following: ethanol, dimethyl sulfoxide and glycerol.
- a method for inhibiting tumors in a host comprising contacting a tumor with an antitumor amount of one or more macrolide compositions of the invention.
- the macrolide compositions of the invention are active for inhibiting a diverse range of tumors including, but not limited to human lung, colon and mammary tumors such as lung carcinoma A549, ileocecal adenocarcinoma HCT-8, and human breast adenocarcinoma cells MDA-MB-231.
- the effectiveness of the compositions of the invention for inhibiting tumors indicates their usefulness for controlling tumors in hosts including mammals and for treating cancerous cachexia.
- an antiviral composition comprising as active ingredient an effective antiviral amount of one or more macrolide compositions of the invention and a non-toxic pharmaceutically acceptable carrier or diluent. While effective amounts may vary, as conditions in which the antiviral- compositions are used vary, a minimal dosage required for activity is generally between 50 to 100 micrograms against 25 to 80 plague forming units of virus cells.
- the macrolide compositions of the invention are active for inhibiting or killing a diverse range of viruses including, but not limited to, RNA viruses, vesicular stomatitis (herein "VSV") adeno-, corona-, reo- and influenza viruses and the DNA virus, Herpes Simplex - I and II (herein “HSV-I” and “HSV-II”) adeno- and papova- viruses.
- VSV vesicular stomatitis
- HSV Herpes Simplex - I and II
- non-toxic pharmaceutically acceptable carriers or diluents include, but are not limited to, the following: ethanol, dimethyl sulfoxide and glycerol.
- a method for inhibiting viruses in a host comprising contacting viruses with an antiviral amount of one or more macrolide compositions of the invention.
- the effectiveness of the macrolide compositions of the invention for inhibiting viruses indicates their usefulness for controlling viruses and virus related diseases in hosts including mammals.
- an antifungal composition is provided comprising as active ingredient an effective antifungal amount of one or more of the macrolide compositions of the invention and a non-toxic pharmaceutically acceptable carrier or diluent. While effective amounts may vary, as conditions in which the antifungal compositions are used vary, a minimal dosage required for activity is generally between 1 and 10 micrograms/ml against 10 3 /ml fungi, such as Candida albicans for example.
- Useful examples of non-toxic pharmaceutically acceptable carriers or diluents include, but are not limited to, the following: ethanol, dimethyl sulfoxide and glycerol.
- a method for inhibiting fungus in a host comprising contacting fungus with an antifungal amount of one or more macrolide compositions of the invention.
- the effectiveness of the compositions of the invention for inhibiting fungus indicates their usefulness for controlling fungus and fungus related diseases in hosts including mammals.
- macrolide compositions may be useful as agricultural fungicides.
- a process is provided to produce the macrolide compositions of the invention.
- the process comprises the steps of collecting samples of the marine sponge of the genus Theonella, contacting the marine sponge with a suitable organic solvent system to obtain an extract; partitioning said extract by reverse phase chromotography to obtain a number of fractions; and isolating a macrolide composition of the invention from the fractionated extract.
- the suitable organic solvent system is selected from the group of solvents consisting of acetone, ethyl acetate, methanol, toluene, methylene chloride, chloroform, methyl ethyl ketone, ethanol, methyl isobutyl ketone and mixtures thereof.
- Particularly preferred extracting solvents are acetone and ethyl acetate. While those solvents listed above are the presently preferred choices for the solvents useful in accordance with the invention, other suitable solvents may be substituted.
- a suitable solvent system should be capable of extracting a macrolide composition of the invention from other components of the sponge.
- compositions according to the invention are synthesized and/or isolated by various fractionation, synthetic and chromatographic techniques from the extracts obtained. Any suitable fractionation and isolation techniques as known to those skilled in the art may be utilized in accordance with the process of the invention. Suitable isolation techniques include various chromotography techniques such as high pressure liquid chromotography (HPLC) with suitable columns as would be known to those skilled in the art (e.g., an ODS column) eluted with a suitable solvent such as, for example, methanol.
- HPLC high pressure liquid chromotography
- the oil showed strong in vitro antitumor activity against P388 mouse leukemia cells, HCT-8 human colon adenocarcinoma, A549 human lung carcinoma, MDA-MB-231 human breast cancer cells.
- the oil was separated using NS gel (polystyrene gel) and eluting with 20:1 methanol-water.
- Misakinolide is reduced to a dihydro or tetrahydro derivative by catalytic reduction.
- a mixture of a sample of misakinolide and a small amount of catalyst such as Pd/C, Pt/C, or Raney Ni in methanol or ethanol is stirred under hydrogen atmosphere to give hydrogenation products.
- catalyst such as Pd/C, Pt/C, or Raney Ni in methanol or ethanol
- Reduction of misakinolide with LiAlH 4 is carried out in dry ether or THF under usual (standard) conditions to yield a product containing a primary alcohol function formed by reduction of the lactonic carbonyl.
- misakinolide in ethanol is heated with potassium hydroxide. After cooling the mixture is acidified by adding hydrochloric acid solution and concentrated. The residue is partitioned between water and ether (or ethyl acetate). The organic phase is concentrated to give a carboxylic acid. If necessary, the product is purified by HPLC,
- a sample of misakinolide in ethanol is heated with hydrochloric acid or sulfuric acid. After cooling and concentration the residue is partitioned between water and ether (or ethyl acetate). The organic layer is concentrated to give a mixture of several dehydration products and/or other acid-catalyzed reaction products which are separated by HPLC.
- P388 mouse leukemia cells are grown in Dulbecco MEM medium with 10% horse serum, 4mM glutamine, and 20 ⁇ g/ml gentamycin (Biologos, Inc.). Cells are incubated in 10% CO 2 and subcultured 2 times per week.
- composition to each well of a 24-well plate or tube and allow solvent to evaporate to dryness.
- IC 50 concentration is the concentration of composition required to inhibit 50% of cell growth on the plate.
- HCT-8 human colon tumor cells are growth in RPMl 1640 medium (GIBCO).
- A549 human lung carcinoma cells are cultured in Dulbecco medium (Biologos, Inc.). All media are supplemented with 10% fetal bovine serum and contain 50 ⁇ g/ml gentamycin. All human tumor cell lines are incubated at 5% CO 2 at 37° and subcultured once a week.
- MDA-MB-231 human breast adenocarcinoma cell line is also utilized in substantial accordance with the above assay.
- Virus a Both herpes simplex type 1 (HSV-1) and vesicular stomatitis virus (VSV) replicate in the CV-1 cell line.
- CV-1 is a fibroblast-like cell culture derived from primary African green monkey cells.
- cell numbers should be approximately 40-50 x 10 6 cells.
- CV-1 cells have a doubling time of 72 hours based on these numbers.
- MCO is a maintenance medium without phenol red made with 1% 4000 centipose methylcellulose.
- FBS is used at 5% level.
- NRMCO is a maintenance overlay medium without phenol red containing 0.1 mg neutral red dye per ml and 2% 15 centipoise methylcellulose.
- d Incubate plates at 37 C and read the following day.
- Antiviral activity should be observed from two parameters. One is actual reduction in the number of plaques and two is the diminution in plaque diameter.
- Antiviral activity is scored from 0 to +++.
- Candida albicans C. albicans(Ca) is grown on Sabouraud dextrose agar to produce single colonies one of which is used to inoculate Sabouraud dextrose broth. The broth is incubated at 37°C with shaking at 200rpm for 18hrs., the resultant culture is frozen with 10% (v/v) glycerol at -80°C and used as the inoculum for the anti-Candida assay.
- Saccharomyces cerevisiae S. cerevisiae (Sc) is grown in a manner identical to that described for C. albicans except that all incubations are at 30°C.
- Penicillium atrovenetum Spores of P. atrovenetum (Pa) are inoculated onto the surface of V8 agar. Incubation at room temperature for approximately 1 week results in a mature fungal colony. The spores are harvested from this plate by washing with 0.1% (v/v) Triton X-100, spores are then washed with distilled water before freezing at -80°C in the presence of 10% (v/v) glycerol.
- C. albicans or S. cerevisiae are inoculated into melted Sabouraud dextrose agar at 45°C to give a cell density of approximately 1000 cells/mL. Plates are prepared with 10mL of the seeded agar in a 10cm x 10cm petri dish. These plates are stored at 4°C until needed for the assay.
- P. atrovenetum spores are inoculated at 1000 conidia/mL into Tryptic Soy agar at 45°C. The remaining protocol is as described for the other microorganisms.
- Paper discs (6.35mm) are impregnated with the test substance and allowed to dry. They are then placed onto the surface of a test plate prepared as detailed above. Plates are incubated overnight (C albicans
- Two-fold dilutions for the drug are prepared in 50uL volumes of Sabouraud dextrose broth using 96-well microtiter plates. An inoculum of C. alibicans is added in a small volume to give a cell density of approximately 1000 cells/mL. Plates are incubated at 37°C overnight. 10uL of Triphenyl tetrazolium chloride (1% w/v) is then added to each well; a further 2 hour incubation results in a deep coloration of the microorganism. The MIC is the lowest concentration of the drug which has completely inhibited growth.
- the above data reports the in vitro activity of misakinolide for inhibiting tumors, viruses and fungi.
- the above results indicate that the macrolide compositions of the invention are useful for inhibiting tumors, viruses and fungi in hosts and the diseases caused thereby.
- the scope of the present invention is not limited by the description, examples, and suggested uses herein and modifications can be made without departing from the spirit of the invention.
- the macrolide compositions of the present invention have a novel molecular skeleton from which many derivative compounds or series of compounds may be prepared. The present invention contemplates such derivatives as modifications of the present invention and within the scope of the invention.
- compositions described herein may have other useful applications such as, for example, analgesic applications or as starting materials for the preparations of other compositions.
- Therapeutic application of the compositions of the present invention can be accomplished by any suitable therapeutic method and technique as is presently or prospectively known to those skilled in the art.
- the present invention cover the modifications and variations of this invention provided that they come within the scope of the appended claims and their equivalents.
Landscapes
- Chemical & Material Sciences (AREA)
- Organic Chemistry (AREA)
- Health & Medical Sciences (AREA)
- Chemical Kinetics & Catalysis (AREA)
- General Chemical & Material Sciences (AREA)
- Medicinal Chemistry (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Pharmacology & Pharmacy (AREA)
- Life Sciences & Earth Sciences (AREA)
- Animal Behavior & Ethology (AREA)
- General Health & Medical Sciences (AREA)
- Public Health (AREA)
- Veterinary Medicine (AREA)
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
Abstract
Novel macrolide compositions which are useful as antitumor, antiviral and antifungal compositions, a process of producing the compositions and a method for inhibiting tumors, viruses and fungi utilizing the compositions. More particularly, the novel macrolide compositions are organic misakinolide and misakinolide derivative of general formula (I), wherein R1, R2, R3, R4, R5 and R6 are the same or different and are a hydrogen, hydroxy, lower alkoxy, or lower acyloxy group, which are derived from marine sponges genus Theonella.
Description
MISAKINOLIDE COMPOSITIONS AND THEIR DERIVATIVES Field of the Invention
This application is a continuation-in-part of Serial No. 879,134 filed June 26, 1986. This invention relates to new organic compositions which have useful antitumor, antiviral and antifungal activity. Additionally and particularly, this invention relates to new antitumor, antiviral and antifungal misakinolide and derivative compositions derived from marine organisms, i.e., sponges of the genus Theonella and their methods of use.
Background of the Invention
Various tumor related diseases inflict man. Considerable research has been devoted to oncology and antitumor measures. Tumors are common in a variety of mammals and the prevention, control of the growth and regression of tumors in mammals is important to man. The term tumor refers to abnormal masses of new tissue growth which is discordant with the economy of the tissue of origin or of the host's body as a whole.
Tumors inflict mammals and man with a variety of disorders and conditions including various forms of cancer and resultant cancerous cachexia. Cancerous cachexia refers to the symptomatic discomfort that accompanies the infliction of a mammal with a tumor. These symptoms include weakened condition of the inflicted mammal as evidenced by, for example, weight loss. The seriousness of cancer is well know, e.g., cancer is second only to heart
and vascular diseases as a cause of death in man.
While certain methods and chemical compositions have been developed which aid in inhibiting, remitting or controlling the growth of tumors further antitumor methods and chemical compositions are needed.
Viral diseases inflict man, plants, insects, and animals. The prevention and control of viral diseases have important health and economic implications.
Viral diseases contribute to inflictions in humans including common colds, herpes and cancer and the importance of their control is obvious. Also important is control of viral diseases in animals for economic reasons as well as the ability of such animals to become virus reservoirs or carriers which facilitate the spreading of viral diseases to humans. Viral plant diseases have been known to have a disruptive effect on the cultivation of fruit trees, tobacco, and various vegetables. Insect viral diseases are also of interest because of the insects' ability to transfer viral diseases to humans.
Prevention of the growth of fungus and the infections and maladies caused by it to mammals and plants is also of importance to man. The presence of fungus may cause various diseases and infections in man including mycotic disease, e.g., pulmonary candidiasis and pulmonary blastomycosis. Certain yeastlike organisms, e.g., Cryptococcus neoformans, may cause serious infections of the central nervous system. More commonly known fungal infections in
humans and mammals include ringworm, which are fungus infections of hair and nail areas, as well as resistant infections of the skin. Many other fungal infections inflict humans and mammals in the areas of skin, mucous membranes, intestinal tract, vaginal area and lungs.
Plants are also attacked by various fungi. Damage caused by fungus infection to agriculture amounts to billions of dollars annually. Various inorganic and organic fungistats and fungicides have been tried but with limited success. It is of course important for the fungistat or fungicide to kill the fungi but not the plant and to leave no toxic residue on the food of the plant. Various methods have been utilized to combat fungus infection in agriculture including foliage fungicide by which method plants are coated with a preventive weather-resistant fungicide. Seed treatment and soil treatment are methods which require fungicides which are safe for seeds and resist degradation by soil and soil microorganisms. Chemotherapeutants are fungicides which permeate the plant to protect new growth or eliminate infections which have already occurred within the plant. Agricultural fungistats and fungicides and their application must also meet very stringent requirements and regulations, which have been promulgated, for example, in the United States.
Considerable research and resources have been devoted to oncology and antitumor measures including chemotherapy; antiviral measures; and combating fungal infections in both mammals and
plants. While various antitumor, antiviral or antifungal agents and methods have been developed which aid in inhibiting tumors, viruses and the spread of fungus, respectively, additional methods and chemical agents are needed.
A potential source for antiviral compositions is marine plant and animal life and of particular interest are marine sponges. It has now been found that certain organic compounds derived from extracts of sponges of the genus Theonella, possess useful antitumor, antiviral and antifungal activity.
Some compounds of interest have been previously isolated from marine sponges and alga. These compounds named swinholide-A and scytophycin A and B have been reported by S. Carmely and Y.
Kashman, "Structure of Swinholide-A, a new macrolide from the Marine Sponge Theonella Swinhoei," Tetrahedron Letters, 26, 511-514 (1985) and R.E. Moore, G.M.L. Patterson, J.S. Mynderse, and J. Barchi, Jr. "Toxins from cyanophytes belonging to the Scytohemataceae" Pure and Applied Chemistry, 58 , No. 2,263-271 (1986), respectively. The entire disclosures of these references is hereby incorporated herein by reference. Thus, marine sponges and other marine life can be a source of useful raw materials for man.
Summary of the Invention
It is therefore an object of the invention
to provide novel compositions which are useful as antitumor, antiviral and antifungal agents and a process for producing such novel compositions.
It is an additional object of the invention to provide a method for inhibiting tumors, viruses and fungus growth and resultant infection and disease utilizing novel antitumor, antiviral and antifungal compositions.
Additional objects and advantages of the invention will be set forth, in part, in the description which follows and in part will be obvious from this description, or may be learned by the practice of the invention. The objects and advantages of the invention are realized and obtained by means of the compositions, processes, methods, and the combinations particularly pointed out in the appended claims.
To achieve the objects in accordance with the purposes of the invention, as embodied and fully described here, the invention comprises compositions of the general formula (I):
wherein R1, R2, R3 R4 R5 and R6 are the same or different and are a hydrogen, hydroxy, lower alkoxy, or lower acyloxy group. The invention also comprises reduced derivatives of these compositions wherein at least one of the double bonds of the composition described above according to formula I is reduced.
The invention also comprises compositions formed by the treatment of any of the compositions of formula I and their reduced derivatives with oxidizing reagents, such as Jones reagent or pyridinium chlorochromate (PCC); with alkaline
solutions; and with acidic solution.
All of the above-described compositions according to formula I, as well as, their reduced derivatives and derivatives formed by treatment with Jones reagent , alkal ine or acid soluti ons will col lectively be referred to, hereinafter, as "macrolide compositions of the invention."
In preferred embodiments of the invention, the composition is substantially pure.
In preferred embodiment of the invention the lower alkoxy and acyloxy groups have from 1 to 5 carbon atoms. In further preferred embodiments of the invention, the invention comprises dimeric compositions of formulae II and ill:
As embodied and fully described herein, the invention also comprises an antitumor, antiviral or
antifungal composition comprising, as active ingredient, an effective antitumor, antiviral or antifungal amount, respectively, of one or more macrolide compositions of the invention and a non-toxic pharmaceutically acceptable carrier or diluent.
As embodied and fully described herein, the invention also comprises a process to produce the macrolide compositions of the invention. The process comprises the steps of collecting marine sponge of the genus Theonella, contacting the marine sponge with a suitable organic solvent system to obtain an extract; fractionating the extract; and isolating a macrolide composition of the invention from the fractionated extract. As embodied and fully described herein, the invention further comprises a method for inhibiting tumors comprising contacting tumor cells with an effective antitumor amount of one or more macrolide compositions of the invention. As embodied and fully described herein, the invention further comprises method for inhibiting viruses comprising contacting viruses with an effective antiviral amount of one or more macrolide compositions of the invention. As embodied and fully described herein, the invention further comprises a method for inhibiting the growth of or killing fungi comprising contacting fungi with an effective antifungal amount of one or more macrolide compositions of the invention. It is to be understood that both the fore-
going general and the following detailed description are exemplary and explanatory only and are not intended to be restrictive of the invention as claimed,
DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS OF THE INVENTION
Reference will now be made in detail to present preferred embodiments of the invention, examples of which are illustrated in the following example section.
In accordance with the invention novel compositions are provided to achieve the objects in accordance with the purposes of the invention, as embodied and fully described herein, the invention comprises macrolide compositions of the general form
wherein R1 R2, R3 R4 R5 and R6 are the
same or different and are a hydrogen, hydroxy, lower alkoxy, or lower acyloxy group. The invention also comprises reduced derivatives of these compositions wherein at least one of the doubl'e bonds of the composition described above according to formula I is reduced.
The invention also comprises compositions formed by the treatment of any of the compositions of formula I and their reduced derivatives with oxidizing agents, such as Jones reagent or PCC; with alkaline solutions; and with acid solutions. All of th.e above-described compositions are referred to herein as "macrolide compositions of the invention."
In preferred embodiments of the invention, the composition is substantially pure.
In preferred embodiments of the invention the lower alkoxy or acyloxy groups have from 1 to 5 carbon atoms. In more preferred embodiments of the invention, the invention comprises compositions of the formulae II and III:
In accordance with the invention, an antitumor composition is provided comprising as active ingredient an effective antitumor amount of one or more of the macrolide compositions of the invention and a non-toxic pharmaceutically acceptable carrier or diluent. While effective amount?, may vary, as conditions in which the antitumor compositions are used vary, a minimal dosage required for activity is generally between 0.01 and 10
5 micrograms against 10 tumor cells, useful examples of non-toxic pharmaceutically acceptable carriers or diluents include, but are not limited to, the following: ethanol, dimethyl sulfoxide and glycerol.
In accordance with the invention, a method for inhibiting tumors in a host is provided comprising contacting a tumor with an antitumor amount of one or more macrolide compositions of the invention. The macrolide compositions of the invention are active for inhibiting a diverse range of tumors including, but not limited to human lung, colon and mammary tumors such as lung carcinoma A549, ileocecal adenocarcinoma HCT-8, and human breast adenocarcinoma cells MDA-MB-231. The effectiveness of the compositions of the invention for inhibiting tumors indicates their usefulness for controlling tumors in hosts including mammals and for treating cancerous cachexia.
In accordance with the invention, an antiviral composition is provided comprising as active ingredient an effective antiviral amount of
one or more macrolide compositions of the invention and a non-toxic pharmaceutically acceptable carrier or diluent. While effective amounts may vary, as conditions in which the antiviral- compositions are used vary, a minimal dosage required for activity is generally between 50 to 100 micrograms against 25 to 80 plague forming units of virus cells. The macrolide compositions of the invention are active for inhibiting or killing a diverse range of viruses including, but not limited to, RNA viruses, vesicular stomatitis (herein "VSV") adeno-, corona-, reo- and influenza viruses and the DNA virus, Herpes Simplex - I and II (herein "HSV-I" and "HSV-II") adeno- and papova- viruses. Useful examples of non-toxic pharmaceutically acceptable carriers or diluents include, but are not limited to, the following: ethanol, dimethyl sulfoxide and glycerol.
In accordance with the invention, a method for inhibiting viruses in a host is provided comprising contacting viruses with an antiviral amount of one or more macrolide compositions of the invention. The effectiveness of the macrolide compositions of the invention for inhibiting viruses indicates their usefulness for controlling viruses and virus related diseases in hosts including mammals. In accordance with the invention, an antifungal composition is provided comprising as active ingredient an effective antifungal amount of one or more of the macrolide compositions of the invention and a non-toxic pharmaceutically acceptable carrier or diluent. While effective amounts may
vary, as conditions in which the antifungal compositions are used vary, a minimal dosage required for activity is generally between 1 and 10 micrograms/ml against 103/ml fungi, such as Candida albicans for example. Useful examples of non-toxic pharmaceutically acceptable carriers or diluents include, but are not limited to, the following: ethanol, dimethyl sulfoxide and glycerol.
In accordance with the invention, a method for inhibiting fungus in a host is provided comprising contacting fungus with an antifungal amount of one or more macrolide compositions of the invention. The effectiveness of the compositions of the invention for inhibiting fungus indicates their usefulness for controlling fungus and fungus related diseases in hosts including mammals. Further, such macrolide compositions may be useful as agricultural fungicides.
In accordance with the invention, a process is provided to produce the macrolide compositions of the invention. The process comprises the steps of collecting samples of the marine sponge of the genus Theonella, contacting the marine sponge with a suitable organic solvent system to obtain an extract; partitioning said extract by reverse phase chromotography to obtain a number of fractions; and isolating a macrolide composition of the invention from the fractionated extract.
In preferred embodiments of the invention the suitable organic solvent system is selected from the group of solvents consisting of acetone, ethyl
acetate, methanol, toluene, methylene chloride, chloroform, methyl ethyl ketone, ethanol, methyl isobutyl ketone and mixtures thereof. Particularly preferred extracting solvents are acetone and ethyl acetate. While those solvents listed above are the presently preferred choices for the solvents useful in accordance with the invention, other suitable solvents may be substituted. A suitable solvent system should be capable of extracting a macrolide composition of the invention from other components of the sponge. Different ratios of solvents and any combination may be used in the solvent system of the invention as would be known to those skilled in the art. Compositions according to the invention are synthesized and/or isolated by various fractionation, synthetic and chromatographic techniques from the extracts obtained. Any suitable fractionation and isolation techniques as known to those skilled in the art may be utilized in accordance with the process of the invention. Suitable isolation techniques include various chromotography techniques such as high pressure liquid chromotography (HPLC) with suitable columns as would be known to those skilled in the art (e.g., an ODS column) eluted with a suitable solvent such as, for example, methanol.
A more detailed description and explanation of a preferred embodiment of the process of the invention to produce a macrolide composition of the invention is provided in the examples section.
It is therefore apparent that the macrolide compositions of the invention, the process for producing the macrolide compositions of the invention and the methods for utilizing the macrolide compositions of the invention to inhibit tumors, viruses and fungus growth fulfill the objects of the invention.
EXAMPLES
The invention will now be illustrated by examples. The examples are not intended to be limiting of the scope of the present invention. In conjunction with the detailed and general description above, the examples provide further understanding of the present invention and outline a process for producing compositions of the invention.
The following examples represent preferred embodiments of the compositions, processes and methods of the invention for satisfying the stated objects of the invention. The starting materials and reagents in the examples whose methods of preparation are not indicated are commercially available from sources known to the art such as chemical supply houses .
EXAMPLE 1
Preparation of Misak inolide ( 1 ) and der ivatives thereof ( 2 - 4 )
A sample (320 g) of marine sponge of the genus Theonella, collected at Maedamisaki, Okinawa, was extracted by steeping in acetone (700 mL) for 48 hours. The extract was concentrated to give an aqueous suspension which on further extraction with ethyl acetate gave 0.5g of ethyl acetate soluble oil. The oil showed strong in vitro antitumor activity against P388 mouse leukemia cells, HCT-8 human colon adenocarcinoma, A549 human lung carcinoma, MDA-MB-231 human breast cancer cells. The oil was separated using NS gel (polystyrene gel) and eluting with 20:1 methanol-water. Active fractions (185 mg) were combined and further separated over silica gel by eluting first with ethyl acetate and
then with 5:1 methylene chloride-methanol. The latter eluate (64 mg) was purified by running on HPLC (ODS column, methanol) to give 24 mg of misakinolide as colorless oil, [ α -21.4° (c 5.6,
CHCl3). Misakinolide gave the following spectral data: UV (MeOH) λmax 220nm (∈ 5000); IR (film) 3440, 2960, 2935, 2820, 1680, 1635, 1455, 1380, 1260, 1195, 1150, 1125, 1080, 985, 970, 925, 850, 750, and 700 cm-1; 1H NMR (C6D6)δ 7.30 (1H, m), 5.69 (1H, br d, J=7.9 Hz), 5.60 (1H, m), 5.55 (1H, br d, J=8 Hz), 4.71 (1H, br d, J=9 Hz), 4.23 (1H, m), 4.18 (1H, m), 4.03 (1H, m), 3.98 (1H, m), 3.88 (1H, m), 3.67 (1H, m), 3.53 (1H, m), 3.34 (3H, s), 3.33 (1H, m), 3.29 (1H, m), 3.15 (3H, s), 1.97 (3H, br s), 1.20 (3H, d, J=4.7 Hz), 1.10 (3H, d, J=5.0 Hz), 1.04 (3H, d, J=4.8 Hz), 1.02 (3H, d, J=5.1 Hz), and 0.95 (3H, d, J=5.0 Hz); 13C NMR (C6D6 171.04 (s), 142.41 (d), 130.55 (d), 129.00 (s), 123.57 (d), 77.74 (d), 76.90 (d), 76.29 (d), 74.85 (d), 73.59 (d), 71.16 (d), 70.74 (d), 68.10 (d), 66.83 (d), 64.68 (d), 64.29 (d), 56.75 (q), 54.99 (q), 41.63 (t), 41.62 (d), 41.12 (d), 39.18 (t), 39.00 (t), 38.20 (t), 37.70 (d), 35.69 (t), 35.23 (t), 33.40 (d), 31.51 (t), 28.55 (t), 24.58 (t), 21.62 (q), 18.20 (q), 12.90 (q), 9.60 (q), 9.46 (q), and 9.46 (q).
Examples 2-4 Treatment of composition (1) with acetone and p-TsOH gives an acetonide ( 2 ). Treatment of acetonide (2) with p-bromobenzoyl chloride and pyridine gives a tribenzoate (3) and tetrabenzoate (4).
Example 5
(1) Catalytic reduction
Misakinolide is reduced to a dihydro or tetrahydro derivative by catalytic reduction. A mixture of a sample of misakinolide and a small amount of catalyst such as Pd/C, Pt/C, or Raney Ni in methanol or ethanol is stirred under hydrogen atmosphere to give hydrogenation products. [2] Reduction with lithium aluminum hydride (LiAlH4)
Reduction of misakinolide with LiAlH4 is carried out in dry ether or THF under usual (standard) conditions to yield a product containing a primary alcohol function formed by reduction of the lactonic carbonyl.
Example 6
(1) Oxidation with Jones reagent
To a stirred solution of misakinolide in acetone or in ether is added Jones reagent prepared in acetone (see Bowers, Halsall, Jones, and Lemin, j.
Chem. Soc., 2548 (1953)) or in ether (see Brown, Garg, and Liu, J. Org. Chem., 36, 387 (1971)) at room temperature. Depending on the amount of the reagent employed and duration of the reaction, products (ketones) are varied and are generally a mixture of several compounds unless the reaction is carried out with an excess of the reagent for a longer period.
Example 7
A sample of misakinolide in ethanol is heated with potassium hydroxide. After cooling the mixture is acidified by adding hydrochloric acid solution and concentrated. The residue is partitioned between water and ether (or ethyl acetate). The organic phase is concentrated to give a carboxylic acid. If necessary, the product is purified by HPLC,
Example 8
A sample of misakinolide in ethanol is heated with hydrochloric acid or sulfuric acid. After cooling and concentration the residue is partitioned between water and ether (or ethyl acetate). The organic layer is concentrated to give a mixture of several dehydration products and/or other acid-catalyzed reaction products which are separated by HPLC.
ANTITUMOR ACTIVITIES OF THE COMPOUNDS OF THE INVENTION
The following assay method was utilized to illustrate the antitumor effectiveness of the compositions of Formulae I and II corresponding to misakinolide of example 1. P388 MOUSE LEUKEMIA CELL ASSAY Maintenance of Cell Line
P388 mouse leukemia cells are grown in
Dulbecco MEM medium with 10% horse serum, 4mM glutamine, and 20μg/ml gentamycin (Biologos, Inc.). Cells are incubated in 10% CO2 and subcultured 2 times per week.
PROCEDURE
1. Add composition to each well of a 24-well plate or tube and allow solvent to evaporate to dryness.
2. Add 2ml (1.2 x 105) cells to each well or tube and mix.
3. Incubate in 10% CO2 at 37° for 48 hours.
4. Read plates with an inverted microscope, scoring activity from 1+ to 4+ as follows: ND (not detectable), >90%; 1+, 75-90%; 2+, 50-74%; 3+, 25-49%; 4+, <25% of control growth. Alternatively, the activity may be designated as IC50 concentration which is the concentration of composition required to inhibit 50% of cell growth on the plate.
Cell counts are performed on each tube and results are reported as percent of control.
HUMAN TUMOR CELL LINE ASSAY
Maintenance of Cell Line HCT-8 human colon tumor cells are growth in RPMl 1640 medium (GIBCO). A549 human lung carcinoma
cells are cultured in Dulbecco medium (Biologos, Inc.). All media are supplemented with 10% fetal bovine serum and contain 50 μg/ml gentamycin. All human tumor cell lines are incubated at 5% CO2 at 37° and subcultured once a week.
PROCEDURE
1. Seed 1ml cell (5000 HCT-8, 8000 A549, 12000 MCF-7) in each well of a 24-well plate.
2. incubate in a CO2-incubator for 48 hours.
3. Add composition to each well and incubate for an additional 120 hours.
4. Discard medium and stain with methylene blue (HCT-8) or crystal violet (A549 and MCF-7).
5. Compare cell density of drug-treated well with that of the control (no drug added) as follows: ND (not detectable), >90%; 1+, 75-90%; 2+, 50-74%; 3+ , 25-49%, 4+,<25% of control growth.
Positive control - Vinblastine or Vincristine in aqueous solution.
Final Cone. of Vinblastine or Vincristine control (use 2 μl/assay)
Solution Cone. Amt added Final cone, in test
5 mg/ml 2 ul 5 μg/ml
1 mg/ml 2 μl 1 μg/ml
0.1 mg/ml 2 μl 0.1 μg/ml
0.05 mg/ml 2 μl 0.05 μg/ml
MDA-MB-231 human breast adenocarcinoma cell line is also utilized in substantial accordance with the above assay.
The results of the above assays are summarized in Table 1.
ANTIVIRAL ACTIVITIES OF THE COMPOSITIONS OF THE INVENTION
The following assay method was utilized to demonstrate the in vitro antiviral effectiveness of compositions of the invention as reported in Table 2,
Antiviral Disc Assay for HSV-1 and VSV
A. Maintenance of Cell cultures
1. Virus a. Both herpes simplex type 1 (HSV-1) and vesicular stomatitis virus (VSV) replicate in the CV-1 cell line. CV-1 is a fibroblast-like cell culture derived from primary African green monkey cells.
2. Growth of CV-1 Cells
2 a. Seed 150cm tissue culture flasks each with 10 x 106 CV-1 cells in 40
ml of EMEM with 10% FBS (growth medium
b. Seven days after seeding the flasks cell numbers should be approximately 40-50 x 106 cells. CV-1 cells have a doubling time of 72 hours based on these numbers.
3. Trypsinization a. Aseptically remove the medium. b. Rinse cell sheet with 10 ml of Ca++ and Mg free Dulbecco's phosphate buffered saline or Pucks G saline at least twice. c. Add 1.5 to 2.0 ml of trypsin -EDTA mixture. d. Incubate flask at room temperature or at 37°C with occasional rocking until the cells detach from the flask (about 15-30 min). Cells maintained on calf serum detach from the plastic at a faster rate than those held on fetal bovine serum (FBS). e. Shake flask. f. Add 10 ml EMEM growth medium and break up cell clumps with pipetting. g. Count cells.
B. Preparation of plates for viral assays 1. Cell Concentration a. Dilute the cells with EMEM to 4 x 105
cells/ml.
Seed 24 well trays with 0.5 ml per well. Cell concentration per well is 2 x 105 cells. c. incubate at 37°C with 5% CO2. d. The wells can be uses over the next several days beginning the day after seeding (preferably 2, 3, or 4).
C. Assay of HSV-1 and VSV in CV-1 cells 1. Infection of CV-1 cells in plates with virus a. Remove medium from wells. b. Infect well with at least 25 and no more than 80 plaque forming units (PFU) of virus. c. Incubate infected cells at 37° C for
1.5 hours. d. Pour off supernatant at end of incubation period. e. Add 0.5 ml of methylcellulose overlay medium (MCO).
1) MCO is a maintenance medium without phenol red made with 1% 4000 centipose methylcellulose. FBS is used at 5% level.
2. Drug Evaluation
a. For drug evaluation wet filter paper discs (6 mm diameter) with approximately 0.02 ml of marine extract
or test compound.
1) Allow solvent to evaporate for 20 to 30 minutes at room temperature.
2) Place discs in the well containing CV-1 cells, virus, and MCO. b. Incubate tissue culture plates for 48 hours at 37° C. c. After 48 hours place 0.5 ml NRMCO on each well.
1) . NRMCO is a maintenance overlay medium without phenol red containing 0.1 mg neutral red dye per ml and 2% 15 centipoise methylcellulose. d. Incubate plates at 37 C and read the following day.
1) Antiviral activity should be observed from two parameters. One is actual reduction in the number of plaques and two is the diminution in plaque diameter.
3. Scoring Drug Activity
a. Antiviral activity is scored from 0 to +++.
+++ = complete inhibition of plaque formation ++ = partial inhibition + = partial inhibition 0 = no protection
b. Cytotoxicity
0 = no visual or microscopic cytotoxicity
16 = Complete cell destruction
8, 10, 12, 14 = partial cytotoxicity
ANTIFUNGAL ACTIVITIES OF THE COMPOSITIONS OF THE INVENTION
The following assay method was utilized to demonstrate the in vitro antifungal effectiveness of compositions of the invention as reported in table 3.
Preparation of inocula
Candida albicans: C. albicans(Ca) is grown on Sabouraud dextrose agar to produce single colonies one of which is used to inoculate Sabouraud dextrose broth. The broth is incubated at 37°C with shaking at 200rpm for 18hrs., the resultant culture is frozen with 10% (v/v) glycerol at -80°C and used as the inoculum for the anti-Candida assay.
Saccharomyces cerevisiae: S. cerevisiae (Sc) is grown in a manner identical to that described for C. albicans except that all incubations are at 30°C.
Penicillium atrovenetum: Spores of P. atrovenetum (Pa) are inoculated onto the surface of V8 agar. Incubation at room temperature for approximately 1 week results in a mature fungal colony. The spores are harvested from this plate by washing with 0.1% (v/v) Triton X-100, spores are then washed with
distilled water before freezing at -80°C in the presence of 10% (v/v) glycerol.
Assay protocols
1. Disc diffusion assay
C. albicans or S. cerevisiae are inoculated into melted Sabouraud dextrose agar at 45°C to give a cell density of approximately 1000 cells/mL. Plates are prepared with 10mL of the seeded agar in a 10cm x 10cm petri dish. These plates are stored at 4°C until needed for the assay.
P. atrovenetum spores are inoculated at 1000 conidia/mL into Tryptic Soy agar at 45°C. The remaining protocol is as described for the other microorganisms.
Paper discs (6.35mm) are impregnated with the test substance and allowed to dry. They are then placed onto the surface of a test plate prepared as detailed above. Plates are incubated overnight (C albicans
37°C; S. cerevisiae 30°C; P. atrovenetum 25°C) after which time the zones of growth inhibition can be read, these are expressed as the diameter of the zone in millimeters.
2. Minimum inhibitory concentration (MIC)
Two-fold dilutions for the drug are prepared in 50uL volumes of Sabouraud dextrose broth using 96-well microtiter plates. An inoculum of C. alibicans is added in a small volume to give a cell density of
approximately 1000 cells/mL. Plates are incubated at 37°C overnight. 10uL of Triphenyl tetrazolium chloride (1% w/v) is then added to each well; a further 2 hour incubation results in a deep coloration of the microorganism. The MIC is the lowest concentration of the drug which has completely inhibited growth.
Table 1
The above data reports the in vitro activity of misakinolide for inhibiting tumors, viruses and fungi. The above results indicate that the macrolide compositions of the invention are useful for inhibiting tumors, viruses and fungi in hosts and the diseases caused thereby.
The scope of the present invention is not limited by the description, examples, and suggested uses herein and modifications can be made without departing from the spirit of the invention. The macrolide compositions of the present invention have a novel molecular skeleton from which many derivative compounds or series of compounds may be prepared. The present invention contemplates such derivatives as modifications of the present invention and within the scope of the invention. For example, it may be noted that other derivatives of the compounds of example 1 such as halogen or alkyl substituted derivatives may be prepared that may possess antitumor, antiviral or antifungal activity analogous to those preferred embodiments described above. Further, the compositions described herein may have other useful applications such as, for example, analgesic applications or as starting materials for the preparations of other compositions. Therapeutic application of the compositions of the present invention can be accomplished by any suitable therapeutic method and technique as is presently or prospectively known to those skilled in the art. Thus, it is intended that the present invention cover the modifications and variations of this invention provided that they come within the scope of the appended claims and their equivalents.
Claims
1. A composition according the formula:
2. A composition according to claim 1 wherein at least one double bond is reduced.
3. A derivative of a composition according to claim 1 which has been oxidized, treated with an alkaline solution or treated with an acid solution.
4. A derivative of a composition according to claim 2 which has been further oxidized, treated with an alkaline solution, or treated with an acid solution.
5. The composition according to claim 1 wherein said composition is substantiaally pure.
6. An antitumor composition comprising, as active ingredient, an effective antitumor amount of one or more of the compositions of claim 1 and a non-toxic pharmaceutically acceptable carrier or diluent.
7. A method for inhibiting tumors in a host comprising contacting a tumor with an effective antitumor amount of one or more compositions of claim
1.
8. A method of inhibiting tumors in a host comprising contacting a tumor with an effective antitumor amount of one or more compositions of claim 3.
9. A method for inhibiting tumors in a mammalian host comprising contacting a tumor with an effective antitumor amount of one or more compositions of claim 1.
10. An antiviral composition comprising, as active ingredient, an effective antiviral amount of one or more of the compositions of claim 1 and a non-toxic pharmaceutically acceptable carrier or diluent.
11. A method for inhibiting viruses comprising contacting viruses with an effective antiviral amount of one or more compositions of claim I-
12. A method of inhibiting viruses in a host comprising contacting viruses with an effective antiviral amount of one or more compositions of claim 1.
13. A method for inhibiting viruses in a mammalian host comprising contacting viruses cells with an effective antiviral amount of one or more compositions of claim 1.
14. An antifungal composition comprising, as active ingredient, an effective antifungal amount of one or more of the compositions of claim 1 and a non-toxic pharmaceutically acceptable carrier or diluent.
15. A method for inhibiting fungus growth comprising contacting fungus with an effective antifungal amount of one or more compositions of claim 1.
16. A method of inhibiting fungus growth in a host comprising contacting fungus with an effective antifungal amount of one or more compositions of claim 3.
17. A method for inhibiting fungus growth in a mammalian host comprising contacting fungus with an effective antifungal amount of one or more compositions of claim 1.
18. A process to produce a composition according to claim 1 comprising the steps of: collecting marine sponge genus Theonella; contacting said sponge with a suitable organic solvent system; obtaining a solvent extract from the sponge; fractionating the extract; and isolating a composition according to claim 1 from the fractionated extract.
19. A therapeutic method for treating cancerous cachexia caused by the presence of a tumor in a host comprising contacting cells of said tumor with an effective antitumor amount of a composition according to claim 1.
20. A therapeutic method for treating cancerous cachexia caused by the presence of a tumor in a host comprising contacting cells of said tumor with an effective antitumor amount of a composition according to claim 3.
21. A composition of the formula:
22. A composition of the formula
Applications Claiming Priority (4)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
US87913486A | 1986-06-26 | 1986-06-26 | |
US879,134 | 1986-06-26 | ||
US07/051,127 US4859782A (en) | 1986-06-26 | 1987-05-18 | Misakinolide compositions and their derivatives |
US051,127 | 1987-05-18 |
Publications (1)
Publication Number | Publication Date |
---|---|
WO1988000195A1 true WO1988000195A1 (en) | 1988-01-14 |
Family
ID=26729098
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
PCT/US1987/001565 WO1988000195A1 (en) | 1986-06-26 | 1987-06-25 | Misakinolide compositions and their derivatives |
Country Status (4)
Country | Link |
---|---|
US (1) | US4859782A (en) |
EP (1) | EP0270660A1 (en) |
JP (1) | JPH01500517A (en) |
WO (1) | WO1988000195A1 (en) |
Cited By (5)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
EP0299713A2 (en) * | 1987-07-17 | 1989-01-18 | Harbor Branch Oceanographic Institution, Inc. | Antiviral, antitumor and antifungal compositions |
US4859782A (en) * | 1986-06-26 | 1989-08-22 | Harbor Branch Oceanographic Institution, Inc. | Misakinolide compositions and their derivatives |
EP0472005A1 (en) * | 1990-07-27 | 1992-02-26 | Bristol-Myers Squibb Company | Antiviral antibiotic BU-4224V |
ES2276629A1 (en) * | 2005-12-15 | 2007-06-16 | Pharma Mar, S.A. | Antitumour compounds |
CN102083840A (en) * | 2008-07-03 | 2011-06-01 | 马尔药品公司 | Antitumoral macrolides |
Families Citing this family (7)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
AU2768592A (en) * | 1991-09-30 | 1993-05-03 | University Of Hawaii | New scytophycin compounds, compositions and methods for their production and use |
US5684036A (en) * | 1994-02-18 | 1997-11-04 | Harbor Branch Oceanographic Institution, Inc. | Cytotoxic macrolides and methods of use |
US5478861A (en) * | 1994-02-18 | 1995-12-26 | Harbor Branch Oceanographic Institution, Inc. | Cytotoxic macrolides and methods of use |
GB9814640D0 (en) * | 1998-07-06 | 1998-09-02 | Fujisawa Pharmaceutical Co | New use |
US20090098078A1 (en) * | 2004-12-23 | 2009-04-16 | Kelvin Brian Dickinson | Water-In-Oil Microemulsions for Hair Treatment |
EP1991270A4 (en) * | 2005-12-22 | 2009-12-02 | Anaborex Inc | Compositions and methods for prevention and treatment of cachexia |
US20090221518A1 (en) * | 2007-11-18 | 2009-09-03 | Anaborex, Inc. | Compositions and Methods for Treatment of Exudative Serous Effusion |
Family Cites Families (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US4859782A (en) * | 1986-06-26 | 1989-08-22 | Harbor Branch Oceanographic Institution, Inc. | Misakinolide compositions and their derivatives |
-
1987
- 1987-05-18 US US07/051,127 patent/US4859782A/en not_active Expired - Fee Related
- 1987-06-25 EP EP87904477A patent/EP0270660A1/en not_active Withdrawn
- 1987-06-25 WO PCT/US1987/001565 patent/WO1988000195A1/en not_active Application Discontinuation
- 1987-06-25 JP JP62504128A patent/JPH01500517A/en active Pending
Non-Patent Citations (1)
Title |
---|
Tetrahedron Letters, Volume 26, NO. 4, 1985, Pergamon Press Ltd, (Oxford, GB), S. CARMELY et al.: "Structure of Swinholide-A, a New Macrolide from the Marine Sponge Theonella Swinhoei", pages 511-514 see page 511, lines 12,13; page 513, formula cited in the application * |
Cited By (7)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US4859782A (en) * | 1986-06-26 | 1989-08-22 | Harbor Branch Oceanographic Institution, Inc. | Misakinolide compositions and their derivatives |
EP0299713A2 (en) * | 1987-07-17 | 1989-01-18 | Harbor Branch Oceanographic Institution, Inc. | Antiviral, antitumor and antifungal compositions |
EP0299713A3 (en) * | 1987-07-17 | 1990-06-13 | Harbor Branch Oceanographic Institution, Inc. | New antiviral, antitumor and antifungal compositions |
EP0472005A1 (en) * | 1990-07-27 | 1992-02-26 | Bristol-Myers Squibb Company | Antiviral antibiotic BU-4224V |
ES2276629A1 (en) * | 2005-12-15 | 2007-06-16 | Pharma Mar, S.A. | Antitumour compounds |
WO2007068776A1 (en) | 2005-12-15 | 2007-06-21 | Pharma Mar, S.A. | Antitumour compounds |
CN102083840A (en) * | 2008-07-03 | 2011-06-01 | 马尔药品公司 | Antitumoral macrolides |
Also Published As
Publication number | Publication date |
---|---|
EP0270660A1 (en) | 1988-06-15 |
US4859782A (en) | 1989-08-22 |
JPH01500517A (en) | 1989-02-23 |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
US4808590A (en) | Antiviral, antitumor and antifungal compositions and their methods of use | |
CA1334414C (en) | Discorhabdin compositions and their methods of use | |
US4859782A (en) | Misakinolide compositions and their derivatives | |
US4708962A (en) | Antiviral and antitumor cyclohexadienone compositions | |
Encarnacion D et al. | Two new flavones from Calliandra californica | |
SK227192A3 (en) | Cyclohexane a tetrahydropyrane derivatives, methods of their production and pharmaceutical agent containing it | |
US4801606A (en) | Antiviral compositions | |
Hoshino et al. | Queenslandon, a new antifungal compound produced by Chrysosporium queenslandicum: production, isolation and structure elucidation | |
US4755529A (en) | Guaiazulene derivatives and their methods of use | |
EP0272810A2 (en) | Antitumor and antiviral alkaloids | |
AU2005226786A1 (en) | Novel cyclopentenedione antifungal compounds and methods for their use | |
TAKAHASHI et al. | The constituents of Lactarius flavidulus Imai | |
EP0285302A1 (en) | Sesquiterpenoid isocyanide composition and its methods of use | |
US4188392A (en) | Antimicrobial compositions | |
EP0289203B1 (en) | Antitumor and antiviral compounds of marine origin | |
US4879307A (en) | Dioxolane antibacterial compositions | |
EP0304157A1 (en) | Antitumor and antiviral alkaloids | |
US4801607A (en) | Antiviral furanoditerpenoids | |
WO1991012250A1 (en) | Novel antiviral and antitumor terpene hydroquinones and methods of use | |
US5204367A (en) | Novel antiviral and anti-leukemia terpene hydroquinones and methods of use | |
US4870099A (en) | Sulfircin and derivatives thereof as therapeutic agents | |
US4921873A (en) | Antiviral and antitumor cyclohexadienone compositions | |
US5051519A (en) | Novel antiviral and antitumor terpene hydroquinones and methods of use | |
WO1987004708A1 (en) | Novel dioxolane compositions and their method of use | |
CA1257281A (en) | Antiviral and anti-tumor cyclohexadienone compositions |
Legal Events
Date | Code | Title | Description |
---|---|---|---|
AK | Designated states |
Kind code of ref document: A1 Designated state(s): JP |
|
AL | Designated countries for regional patents |
Kind code of ref document: A1 Designated state(s): AT BE CH DE FR GB IT LU NL SE |
|
WWE | Wipo information: entry into national phase |
Ref document number: 1987904477 Country of ref document: EP |
|
WWP | Wipo information: published in national office |
Ref document number: 1987904477 Country of ref document: EP |
|
WWW | Wipo information: withdrawn in national office |
Ref document number: 1987904477 Country of ref document: EP |