PL240096B1 - 4'-O-β-D-(4"-O-Methylglucopyranosyl)-2',5'-dimethoxyflavone and method of preparing 4'-O-β-D- (4"-O-methylglucopyranosyl) -2', 5'-dimetoksyflavone - Google Patents
4'-O-β-D-(4"-O-Methylglucopyranosyl)-2',5'-dimethoxyflavone and method of preparing 4'-O-β-D- (4"-O-methylglucopyranosyl) -2', 5'-dimetoksyflavone Download PDFInfo
- Publication number
- PL240096B1 PL240096B1 PL428743A PL42874319A PL240096B1 PL 240096 B1 PL240096 B1 PL 240096B1 PL 428743 A PL428743 A PL 428743A PL 42874319 A PL42874319 A PL 42874319A PL 240096 B1 PL240096 B1 PL 240096B1
- Authority
- PL
- Poland
- Prior art keywords
- methylglucopyranosyl
- dimethoxyflavone
- organic solvent
- kch
- culture
- Prior art date
Links
- 238000000034 method Methods 0.000 title claims description 14
- 241000751139 Beauveria bassiana Species 0.000 claims abstract description 8
- 239000003960 organic solvent Substances 0.000 claims abstract description 6
- YVAMPWXEJCQAJJ-UHFFFAOYSA-N 2-(2,5-dimethoxyphenyl)chromen-4-one Chemical compound COC1=CC=C(OC)C(C=2OC3=CC=CC=C3C(=O)C=2)=C1 YVAMPWXEJCQAJJ-UHFFFAOYSA-N 0.000 claims description 8
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 claims description 6
- 239000000758 substrate Substances 0.000 claims description 6
- 230000009466 transformation Effects 0.000 claims description 5
- HEDRZPFGACZZDS-UHFFFAOYSA-N Chloroform Chemical compound ClC(Cl)Cl HEDRZPFGACZZDS-UHFFFAOYSA-N 0.000 claims description 4
- 238000004587 chromatography analysis Methods 0.000 claims description 3
- 230000008569 process Effects 0.000 claims description 3
- 241000233866 Fungi Species 0.000 claims description 2
- 239000003480 eluent Substances 0.000 claims description 2
- 239000000203 mixture Substances 0.000 claims description 2
- 239000011541 reaction mixture Substances 0.000 claims description 2
- XEKOWRVHYACXOJ-UHFFFAOYSA-N Ethyl acetate Chemical compound CCOC(C)=O XEKOWRVHYACXOJ-UHFFFAOYSA-N 0.000 abstract description 9
- 230000033444 hydroxylation Effects 0.000 abstract description 3
- 238000005805 hydroxylation reaction Methods 0.000 abstract description 3
- 238000004519 manufacturing process Methods 0.000 abstract description 3
- 238000000605 extraction Methods 0.000 abstract description 2
- 244000005700 microbiome Species 0.000 abstract description 2
- 230000002255 enzymatic effect Effects 0.000 abstract 1
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 abstract 1
- IAZDPXIOMUYVGZ-UHFFFAOYSA-N Dimethylsulphoxide Chemical compound CS(C)=O IAZDPXIOMUYVGZ-UHFFFAOYSA-N 0.000 description 8
- 229930003935 flavonoid Natural products 0.000 description 5
- 150000002215 flavonoids Chemical class 0.000 description 5
- 235000017173 flavonoids Nutrition 0.000 description 5
- 125000000956 methoxy group Chemical group [H]C([H])([H])O* 0.000 description 5
- 229930003944 flavone Natural products 0.000 description 4
- 150000002213 flavones Chemical class 0.000 description 4
- 235000011949 flavones Nutrition 0.000 description 4
- 241000196324 Embryophyta Species 0.000 description 3
- 238000002360 preparation method Methods 0.000 description 3
- YEHDMSUNJUONMW-UHFFFAOYSA-N 2'-methoxyflavone Chemical compound COC1=CC=CC=C1C1=CC(=O)C2=CC=CC=C2O1 YEHDMSUNJUONMW-UHFFFAOYSA-N 0.000 description 2
- 102000004190 Enzymes Human genes 0.000 description 2
- 108090000790 Enzymes Proteins 0.000 description 2
- 101100369992 Homo sapiens TNFSF10 gene Proteins 0.000 description 2
- 241000701024 Human betaherpesvirus 5 Species 0.000 description 2
- 102000046283 TNF-Related Apoptosis-Inducing Ligand Human genes 0.000 description 2
- 108700012411 TNFSF10 Proteins 0.000 description 2
- 101150044134 US28 gene Proteins 0.000 description 2
- 239000000556 agonist Substances 0.000 description 2
- 230000000845 anti-microbial effect Effects 0.000 description 2
- 239000003963 antioxidant agent Substances 0.000 description 2
- 230000003078 antioxidant effect Effects 0.000 description 2
- 235000006708 antioxidants Nutrition 0.000 description 2
- 238000006243 chemical reaction Methods 0.000 description 2
- 230000000694 effects Effects 0.000 description 2
- 229930014626 natural product Natural products 0.000 description 2
- 102000035025 signaling receptors Human genes 0.000 description 2
- 108091005475 signaling receptors Proteins 0.000 description 2
- BWAGQNPYTNMEJP-UHFFFAOYSA-N 2-phenylchromene-4-thione Chemical class O1C2=CC=CC=C2C(=S)C=C1C1=CC=CC=C1 BWAGQNPYTNMEJP-UHFFFAOYSA-N 0.000 description 1
- ZAIANDVQAMEDFL-UHFFFAOYSA-N 3-methoxy-2-phenylchromen-4-one Chemical class O1C2=CC=CC=C2C(=O)C(OC)=C1C1=CC=CC=C1 ZAIANDVQAMEDFL-UHFFFAOYSA-N 0.000 description 1
- 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 1
- 102000000340 Glucosyltransferases Human genes 0.000 description 1
- 108010055629 Glucosyltransferases Proteins 0.000 description 1
- 102000051366 Glycosyltransferases Human genes 0.000 description 1
- 108700023372 Glycosyltransferases Proteins 0.000 description 1
- CSNNHWWHGAXBCP-UHFFFAOYSA-L Magnesium sulfate Chemical compound [Mg+2].[O-][S+2]([O-])([O-])[O-] CSNNHWWHGAXBCP-UHFFFAOYSA-L 0.000 description 1
- 235000002343 Primula veris Nutrition 0.000 description 1
- 244000072254 Primula veris Species 0.000 description 1
- 235000011449 Rosa Nutrition 0.000 description 1
- 101100434907 Rosa hybrid cultivar RhGT1 gene Proteins 0.000 description 1
- 239000013543 active substance Substances 0.000 description 1
- 230000003266 anti-allergic effect Effects 0.000 description 1
- 230000000844 anti-bacterial effect Effects 0.000 description 1
- 229940046836 anti-estrogen Drugs 0.000 description 1
- 230000001833 anti-estrogenic effect Effects 0.000 description 1
- 230000003110 anti-inflammatory effect Effects 0.000 description 1
- 230000000118 anti-neoplastic effect Effects 0.000 description 1
- 230000006907 apoptotic process Effects 0.000 description 1
- 230000008901 benefit Effects 0.000 description 1
- 230000015572 biosynthetic process Effects 0.000 description 1
- 239000003795 chemical substances by application Substances 0.000 description 1
- 231100000433 cytotoxic Toxicity 0.000 description 1
- 230000001472 cytotoxic effect Effects 0.000 description 1
- FMGSKLZLMKYGDP-USOAJAOKSA-N dehydroepiandrosterone Chemical compound C1[C@@H](O)CC[C@]2(C)[C@H]3CC[C@](C)(C(CC4)=O)[C@@H]4[C@@H]3CC=C21 FMGSKLZLMKYGDP-USOAJAOKSA-N 0.000 description 1
- 235000005911 diet Nutrition 0.000 description 1
- 230000037213 diet Effects 0.000 description 1
- 201000010099 disease Diseases 0.000 description 1
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 1
- 239000000328 estrogen antagonist Substances 0.000 description 1
- 239000003172 expectorant agent Substances 0.000 description 1
- 230000003419 expectorant effect Effects 0.000 description 1
- 210000001035 gastrointestinal tract Anatomy 0.000 description 1
- 239000008103 glucose Substances 0.000 description 1
- 230000013595 glycosylation Effects 0.000 description 1
- 238000006206 glycosylation reaction Methods 0.000 description 1
- 108700014210 glycosyltransferase activity proteins Proteins 0.000 description 1
- 230000036541 health Effects 0.000 description 1
- 238000004128 high performance liquid chromatography Methods 0.000 description 1
- 238000000338 in vitro Methods 0.000 description 1
- 239000004615 ingredient Substances 0.000 description 1
- 201000004792 malaria Diseases 0.000 description 1
- 230000007246 mechanism Effects 0.000 description 1
- 230000001404 mediated effect Effects 0.000 description 1
- 230000000813 microbial effect Effects 0.000 description 1
- 230000008520 organization Effects 0.000 description 1
- 230000000144 pharmacologic effect Effects 0.000 description 1
- 239000002904 solvent Substances 0.000 description 1
- 230000003595 spectral effect Effects 0.000 description 1
- 238000003786 synthesis reaction Methods 0.000 description 1
- 230000001225 therapeutic effect Effects 0.000 description 1
Classifications
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
- C12P19/00—Preparation of compounds containing saccharide radicals
- C12P19/44—Preparation of O-glycosides, e.g. glucosides
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07H—SUGARS; DERIVATIVES THEREOF; NUCLEOSIDES; NUCLEOTIDES; NUCLEIC ACIDS
- C07H15/00—Compounds containing hydrocarbon or substituted hydrocarbon radicals directly attached to hetero atoms of saccharide radicals
- C07H15/26—Acyclic or carbocyclic radicals, substituted by hetero rings
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12R—INDEXING SCHEME ASSOCIATED WITH SUBCLASSES C12C - C12Q, RELATING TO MICROORGANISMS
- C12R2001/00—Microorganisms ; Processes using microorganisms
- C12R2001/645—Fungi ; Processes using fungi
Landscapes
- Chemical & Material Sciences (AREA)
- Organic Chemistry (AREA)
- Engineering & Computer Science (AREA)
- Life Sciences & Earth Sciences (AREA)
- Biochemistry (AREA)
- Biotechnology (AREA)
- Genetics & Genomics (AREA)
- General Health & Medical Sciences (AREA)
- Wood Science & Technology (AREA)
- Zoology (AREA)
- Health & Medical Sciences (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Bioinformatics & Cheminformatics (AREA)
- General Engineering & Computer Science (AREA)
- Microbiology (AREA)
- General Chemical & Material Sciences (AREA)
- Molecular Biology (AREA)
- Preparation Of Compounds By Using Micro-Organisms (AREA)
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
Abstract
Wynalazek dotyczy 4'-O-ß-D-(4"-O-metyloglukopiranozylo)-2',5'-dimetoksyflawonu i sposobu wytwarzania 4'-O-ß-D-(4"-O-metyloglukopiranozylo)-2',5'-dimetoksyflawonu o wzorze 2. W wyniku działania układu enzymatycznego zawartego w komórkach szczepu Beauveria bassiana KCh J3.2, następuje hydroksylacja i 4"-O-metyloglikozylacja. Uzyskany w ten sposób produkt wydziela się z wodnej kultury mikroorganizmu, znanym sposobem, przez ekstrakcję rozpuszczalnikiem organicznym niemieszającym się z wodą (octan etylu).The invention relates to 4'-O-ß-D-(4"-O-methylglucopyranosyl)-2',5'-dimethoxyflavone and a method for producing 4'-O-ß-D-(4"-O-methylglucopyranosyl)-2' ,5'-dimethoxyflavone of formula 2. As a result of the action of the enzymatic system contained in the cells of the Beauveria bassiana KCh J3.2 strain, hydroxylation and 4"-O-methylglycosylation occur. The product obtained in this way is isolated from the aqueous culture of the microorganism in a known way, by extraction with an organic solvent immiscible with water (ethyl acetate).
Description
Przedmiotem wynalazku jest sposób wytwarzania 4’-O -β-β-(4”-O-metyloglukopiranozylo)-2’,5’-dimetoksyfl awo nu.The present invention relates to a process for the production of 4'-O -β-β- (4 "-O-methylglucopyranosyl) -2 ', 5'-dimethoxyflawon.
Metoda, według wynalazku może znaleźć zastosowanie w przemyśle farmaceutycznym do wytwarzania składników preparatów poprawiających funkcjonowanie przewodu pokarmowego.The method according to the invention may find application in the pharmaceutical industry for the production of ingredients of preparations improving the functioning of the gastrointestinal tract.
Flawony są syntezowane przez rośliny i stanowią składnik naszej diety o wysokim potencjale antyoksydacyjnym oraz zdolności do modulowania wielu układów enzymatycznych związanych z wieloma chorobami (Mughal E.U., Ayaz M., Hussain Z., Hasan A., Sadiq A., Riaz M., Malik A., Hussain S., Choudhary M.l. 2006. Synthesis and antibacterial activity of substituted flavones, 4-thioflavones and 4-iminoflavones. Bioorg. Med. Chem. 14: 4704-4711). Różne naturalne, półsyntetyczne i syntetyczne flawony zostały scharakteryzowane pod kątem szeregu działań terapeutycznych, takich jak działania przeciwzapalne, przeciwestrogenowe, przeciwdrobnoustrojowe (Cushnie T.P.T., Lamb A.J. 2005. Antimicrobialactivity of flavonoids. Int. J. Antimicrol. Agents 26: 343-35), przeciwalergiczne, przeciwutleniające, przeciwnowotworowe czy też cytotoksyczne (Havsteen B. 1983. Flavonoids, a class of natural products of high pharmacological potency. Biochem. Pharmacol. 32: 1141-1148; Aregawi M., Williams R., Dye C., Cibulskis R., Otten M. 2008. World Malaria 2008, World Health Organization (WHO): Geneva).2’,5’-dimetoksyflawon był wyizolowany z kultury tkankowej rośliny leczniczej Primulaverisz której wykonuje się preparaty wykrztuśne (Jaromir Budzianowski, Maria Morozowska, Maria Wesołowska. Lipophilic flavones of Primula veris L. from field cultivation and in vitro cultures. Phytochemistry 66 (2005) 1033-1039). 2’-metoksyflawon jest aktywnym czynnikiem wywołującym apoptozę indukowaną przez TRAIL w ludzkich białaczkowych komórkach MOLT-4 (Benjawan Wudtiwai, Bungorn Sripanidkulchai, Prachya Kongtawelert, Ratana Banjerdpongchai. Methoxyflavone derivatives modulate the effect of TRAIL-induced apoptosis in human leukemic cell lines. Journal of Hematology & Oncology 2011,4:52).Flavones are synthesized by plants and are part of our diet with high antioxidant potential and the ability to modulate many enzyme systems associated with many diseases (Mughal EU, Ayaz M., Hussain Z., Hasan A., Sadiq A., Riaz M., Malik A., Hussain S., Choudhary MI 2006. Synthesis and antibacterial activity of substituted flavones, 4-thioflavones and 4-iminoflavones. Bioorg. Med. Chem. 14: 4704-4711). Various natural, semi-synthetic and synthetic flavones have been characterized for a number of therapeutic activities, such as anti-inflammatory, anti-estrogen, and antimicrobial activities (Cushnie TPT, Lamb AJ 2005. Antimicrobialactivity of flavonoids. Int. J. Antimicrol. Agents 26: 343-35), antiallergic. , antioxidant, antineoplastic or cytotoxic (Havsteen B. 1983. Flavonoids, a class of natural products of high pharmacological potency. Biochem. Pharmacol. 32: 1141-1148; Aregawi M., Williams R., Dye C., Cibulskis R. , Otten M. 2008. World Malaria 2008, World Health Organization (WHO): Geneva) .2 ', 5'-dimethoxyflavone was isolated from the tissue culture of the medicinal plant Primulaverisz for expectorant preparations (Jaromir Budzianowski, Maria Morozowska, Maria Wesołowska. Lipophilic flavones of Primula veris L. from field cultivation and in vitro cultures. Phytochemistry 66 (2005) 1033-1039). 2'-methoxyflavone is an active agent of TRAIL-induced apoptosis in human MOLT-4 leukemic cells (Benjawan Wudtiwai, Bungorn Sripanidkulchai, Prachya Kongtawelert, Ratana Banjerdpongchai. Methoxyflavone derivatives modulate the effect of TRAIL-induced lines in human leukoptosis. Hematology & Oncology 2011.4: 52).
2’-metoksy-5’-halogenoflawony wykazują aktywność odwrotnych agonistów kluczowego receptora sygnałowego US28 ludzkiego wirusa cytomegalii (Ana Kralj, Mai-Thao Nguyen, NuskaTschammer, Nicolette Ocampo, Quinto Gesiotto, Markus R. Heinrich, Otto Phanstiel, Development of Flavonoid-Based Inverse Agonists of the Key Signaling Receptor US28 of Human Cytomegalovirus. J Med Chem. 2013 Jun 27; 56(12): 5019-32). Obecność jednostki cukrowej ma wpływ na stabilność i rozpuszczalność flawonoidów, a często także wpływa na ich biodostępność i bioaktywność (Wang X. Structure, mechanism and engineering of plant natural product glycosyltransferases. FEBS Lett. 2009; 583(20):3303-9; Wang L, Han W, Xie C, Hou J, Fang Q, Gu J, et al. Comparing the acceptor promiscuity of a Rosa hybridaglucosyltransferase RhGT1 and an engineered microbial glucosyltransferase OleDPSA toward a small flavonoid library. Carbohydr Res. 2013; 368:73-7; Pandey RP, Gurung RB, Parajuli P, Koirala N, Tuoi LT, Sohng JK. Assessing acceptor substrate promiscuity of YjiC-mediated glycosylation toward flavonoids. Carbohydr Res. 2014; 393:26-31).2'-methoxy-5'-halogenoflavones exhibit the activity of inverse agonists of the key US28 signaling receptor of the human cytomegalovirus (Ana Kralj, Mai-Thao Nguyen, NuskaTschammer, Nicolette Ocampo, Quinto Gesiotto, Markus R. Heinrich, Otto Phanstiel, Development of Flavonoid-Based Inverse Agonists of the Key Signaling Receptor US28 of Human Cytomegalovirus. J Med Chem. 2013 Jun 27; 56 (12): 5019-32). The presence of a sugar unit affects the stability and solubility of flavonoids, and often also affects their bioavailability and bioactivity (Wang X. Structure, mechanism and engineering of plant natural product glycosyltransferases. FEBS Lett. 2009; 583 (20): 3303-9; Wang L, Han W, Xie C, Hou J, Fang Q, Gu J, et al. Comparing the acceptor promiscuity of a Rosa hybridaglucosyltransferase RhGT1 and an engineered microbial glucosyltransferase OleDPSA toward a small flavonoid library. Carbohydr Res. 2013; 368: 73- 7; Pandey RP, Gurung RB, Parajuli P, Koirala N, Tuoi LT, Sohng JK. Assessing acceptor substrate promiscuity of YjiC-mediated glycosylation toward flavonoids. Carbohydr Res. 2014; 393: 26-31).
W literaturze nie ma doniesień dotyczących uzyskania 4’-O-e-D-(4”- O-metyloglukopiranozylo)-2’,5’-dimetoksyflawonu przy udziale Beauveria bassiana KCh J3.2.There are no reports in the literature on obtaining 4'-O-e-D- (4 "- O-methylglucopyranosyl) -2", 5'-dimethoxyflavone with Beauveria bassiana KCh J3.2.
Szczep Beauveria bassiana KCh J3.2 był wcześniej ujawniony w literaturze (Kozłowska E, Urbaniak M, Hoc N, Grzeszczuk J, Dymarska M, Stępień Ł, Pląskowska E, Kostrzewa-Susłow E, Janeczko T. (2018) Cascadebiotransformation of dehydroepiandrosterone (DHEA) by Beauveriaspecies. ScientificReports, 8:13449)The Beauveria bassiana KCh J3.2 strain was previously disclosed in the literature (Kozłowska E, Urbaniak M, Hoc N, Grzeszczuk J, Dymarska M, Stępień Ł, Pląskowska E, Kostrzewa-Susłow E, Janeczko T. (2018) Cascadebiotransformation of dehydroepiandrosterone (DHEA ) by Beauveriaspecies. ScientificReports, 8: 13449)
Istota wynalazku polega na tym, że do podłoża odpowiedniego dla grzybów strzępkowych wprowadza się szczep Beauveria bassiana KCh J3.2. Po upływie co najmniej 48 godzin do hodowli wprowadza się substrat, którym jest 2’,5'-dimetoksyflawon wzorze 1, rozpuszczony w rozpuszczalniku organicznym mieszającym się z wodą. T ransformację prowadzi się w temperaturze od 20 do 30 stopni Celsjusza, przy ciągłym wstrząsaniu, co najmniej 1 dobę. Kolejno produkt ekstrahuje się rozpuszczalnikiem organicznym niemieszającym się z wodą i oczyszcza chromatograficznie.The essence of the invention consists in introducing the Beauveria bassiana KCh J3.2 strain into a medium suitable for filamentous fungi. After at least 48 hours, the substrate is introduced into the culture, which is 2 ', 5'-dimethoxyflavone of formula 1, dissolved in a water-miscible organic solvent. The transformation is carried out at a temperature of 20 to 30 degrees Celsius, with continuous shaking, for at least 1 day. The product is subsequently extracted with a water-immiscible organic solvent and purified by chromatography.
W wyniku regioselektywnej hydroksylacji i 4”-O-metyloglikozylacji otrzymuje się 4’-O-e-D-(4”-O -metyloglukopiranozylo)-2’,5’-dimetoksyflawon, a reakcję prowadzi się w wodnej kulturze szczepu Beauveria bassiana KCh J3.2.Regioselective hydroxylation and 4 "-O-methylglycosylation give 4'-O-e-D- (4" -O-methylglucopyranosyl) -2 ", 5'-dimethoxyflavone, and the reaction is carried out in an aqueous culture of Beauveria bassiana KCh J3.2.
Korzystnie jest, gdy stosunek masy dodawanego substratu do objętości hodowli wynosi 0,2 g : 1 L.Preferably, the ratio of the mass of the added substrate to the culture volume is 0.2 g: 1 L.
Korzystnie także jest, gdy proces prowadzi się w temperaturze 25 stopni Celsjusza.It is also preferred that the process is carried out at a temperature of 25 degrees Celsius.
Dodatkowo, korzystnie jest, gdy transformację prowadzi się przez 6 dni.Additionally, it is preferred that the transformation is carried out for 6 days.
PL 240 096 B1PL 240 096 B1
Postępując zgodnie z wynalazkiem, w wyniku działania układu enzymatycznego zawartego w komórkach szczepu Beauveria bassiana KCh J3.2, następuje regioselektywna hydroksylacja i 4 - O-metyloglukozylacja substratu. Uzyskany w ten sposób produkt wydziela się z wodnej kultury mikroorganizmu, znanym sposobem, przez ekstrakcję rozpuszczalnikiem organicznym niemieszającym się z wodą (octan etylu). Zasadniczą zaletą wynalazku jest otrzymanie 4’-O-e-D-(4”-O-metyloglukopiranozylo)-2’,5’-dimetoksyflawonu, z wydajnością izolowaną na poziomie 5% (konwersją według HPLC = 7%),w temperaturze pokojowej i przy pH naturalnym dla szczepu.By following the invention, regioselective hydroxylation and 4 - O-methylglucosylation of the substrate take place as a result of the enzyme system contained in the cells of the strain Beauveria bassiana KCh J3.2. The product obtained in this way is separated from the aqueous culture of the microorganism by a known method by extraction with a water-immiscible organic solvent (ethyl acetate). The main advantage of the invention is the preparation of 4'-OeD- (4 "-O-methylglucopyranosyl) -2 ', 5'-dimethoxyflavone with an isolated yield of 5% (conversion according to HPLC = 7%), at room temperature and natural pH for the strain.
Wynalazek jest bliżej objaśniony na przykładzie wykonania.The invention is explained in more detail using an exemplary embodiment.
P r z y k ł a d. Do kolby Erlenmajera o pojemności 2000 cm3, w której znajduje się 500 cm3 sterylnej pożywki zawierającej 5 g aminobaku i 15 g glukozy, wprowadza się szczep Beauveria bassiana KCh J3.2. Po 72 godzinach jego wzrostu dodaje się 100 mg 2’,5'-dimetoksyflawonu o wzorze 1, rozpuszczonego w 2 cm3 dimetylosuflotlenku (DMSO). T ransformację prowadzi się w 25 stopniach Celsjusza przy ciągłym wstrząsaniu przez 3 dni. Następnie mieszaninę poreakcyjną ekstrahuje się trzykrotnie octanem etylu, osusza bezwodnym siarczanem magnezu i odparowuje rozpuszczalnik. Otrzymany ekstrakt oczyszcza się chromatograficznie, używając jako eluentu mieszaniny chloroform i metanol 9:1.EXAMPLE The Beauveria bassiana KCh J3.2 strain is introduced into a 2000 cc Erlenmajer flask containing 500 cc of sterile medium containing 5 g of aminobac and 15 g of glucose. After 72 hours of growth, 100 mg of 2 ', 5'-dimethoxyflavone I, dissolved in 2 cm 3 of dimethylsulfoxide (DMSO), are added. The transformation is carried out at 25 degrees Celsius with continuous shaking for 3 days. The reaction mixture was then extracted three times with ethyl acetate, dried with anhydrous magnesium sulfate and the solvent was evaporated. The extract obtained is purified by chromatography using a 9: 1 mixture of chloroform and methanol as eluent.
Uzyskany produkt charakteryzuje się następującymi danymi spektralnymi:The obtained product is characterized by the following spectral data:
1H NMR (600 MHz) (DMSO) δ (ppm): 3.00 (t, 1 H, J = 9.2 Hz, H-4”), 3.32-3.37 (m, 1 H, H-2”), 3.39-3.43 (m, 1H, H-5”), 3.46 (s, 3H, C-4”-OCH3), 3.47-3.54 (m, 2H, H-3” and one of H-6”), 3.64-3.69 (m, 1H, one of H-6”), 3.84 (s, 3H, C-5”-OCH3), 3.90 (s, 3H, C-2”-OCH3), 4.79 (t, 1 H, J = 5.1 Hz, C-6”OH), 5.14 (d, 1 H, J = 7.9 Hz, H-1”), 5.32 (d, 1H, J = 5.6 Hz, C-3”-OH), 5.49 (d, 1 H, J = 5.6 Hz, C-2”OH), 6.98 (s, 1H, H-3), 7.00 (s, 1H, H-3’), 7.48 (ddd, 1 H, J = 8.0, 7.1, 1.3 Hz, H-6), 7.56 (s, 1 H, H-6’), 7.79 (dd, 1 H, J = 8.4, 1.3 Hz, H-8), 7.81 (ddd, 1 H, J = 8.5, 7.1, 1.5 Hz, H-7), 8.03 (dd, 1 H, J = 7.9, 1.5 Hz, H-5). 1 H NMR (600 MHz) (DMSO) δ (ppm): 3.00 (t, 1H, J = 9.2 Hz, H-4 "), 3.32-3.37 (m, 1H, H-2"), 3.39- 3.43 (m, 1H, H-5 "), 3.46 (s, 3H, C-4" -OCH3), 3.47-3.54 (m, 2H, H-3 "and one of H-6"), 3.64-3.69 (m, 1H, one of H-6 ”), 3.84 (s, 3H, C-5” -OCH3), 3.90 (s, 3H, C-2 ”-OCH3), 4.79 (t, 1H, J = 5.1 Hz, C-6 ”OH), 5.14 (d, 1H, J = 7.9 Hz, H-1”), 5.32 (d, 1H, J = 5.6 Hz, C-3 ”-OH), 5.49 (d , 1H, J = 5.6 Hz, C-2 ”OH), 6.98 (s, 1H, H-3), 7.00 (s, 1H, H-3 '), 7.48 (ddd, 1H, J = 8.0, 7.1,1.3 Hz, H-6), 7.56 (s, 1H, H-6 '), 7.79 (dd, 1H, J = 8.4, 1.3Hz, H-8), 7.81 (ddd, 1H, J = 8.5, 7.1, 1.5 Hz, H-7), 8.03 (dd, 1H, J = 7.9, 1.5 Hz, H-5).
13 C NMR (151 MHz, DMSO) δ = 56.41 (C-2’-OCHs), 56.69 (C-6’-OCH3), 59.75 (C-4”-OCH3), 60.39 (C-6”), 73.27 (C-2”), 75.96 (C-5”), 76.67 (C-3”), 79.36 (C-4”), 99.48 (C-1”), 101.40 (C-3’), 110.56 (C-3), 111.82 (C-1’), 113.06 (C-6’), 119.60 (C-8), 123.11 (C-4a), 124.66 (C-5), 125.29 (C-6), 134.33 (C-7), 142.81 (C-4’), 150.23 (C-5’), 153.23 (C-2’), 155.78 (C-8a), 160.31 (C-2), 177.12 (C-4). 13 C NMR (151 MHz, DMSO) δ = 56.41 (C-2'-OCHs), 56.69 (C-6'-OCH3), 59.75 (C-4 "-OCH3), 60.39 (C-6"), 73.27 (C-2 ”), 75.96 (C-5”), 76.67 (C-3 ”), 79.36 (C-4”), 99.48 (C-1 ”), 101.40 (C-3 '), 110.56 (C -3), 111.82 (C-1 '), 113.06 (C-6'), 119.60 (C-8), 123.11 (C-4a), 124.66 (C-5), 125.29 (C-6), 134.33 ( C-7), 142.81 (C-4 '), 150.23 (C-5'), 153.23 (C-2 '), 155.78 (C-8a), 160.31 (C-2), 177.12 (C-4).
Claims (5)
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| PL428743A PL240096B1 (en) | 2019-01-31 | 2019-01-31 | 4'-O-β-D-(4"-O-Methylglucopyranosyl)-2',5'-dimethoxyflavone and method of preparing 4'-O-β-D- (4"-O-methylglucopyranosyl) -2', 5'-dimetoksyflavone |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| PL428743A PL240096B1 (en) | 2019-01-31 | 2019-01-31 | 4'-O-β-D-(4"-O-Methylglucopyranosyl)-2',5'-dimethoxyflavone and method of preparing 4'-O-β-D- (4"-O-methylglucopyranosyl) -2', 5'-dimetoksyflavone |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| PL428743A1 PL428743A1 (en) | 2020-08-10 |
| PL240096B1 true PL240096B1 (en) | 2022-02-14 |
Family
ID=71943753
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| PL428743A PL240096B1 (en) | 2019-01-31 | 2019-01-31 | 4'-O-β-D-(4"-O-Methylglucopyranosyl)-2',5'-dimethoxyflavone and method of preparing 4'-O-β-D- (4"-O-methylglucopyranosyl) -2', 5'-dimetoksyflavone |
Country Status (1)
| Country | Link |
|---|---|
| PL (1) | PL240096B1 (en) |
-
2019
- 2019-01-31 PL PL428743A patent/PL240096B1/en unknown
Also Published As
| Publication number | Publication date |
|---|---|
| PL428743A1 (en) | 2020-08-10 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Oglah et al. | Synthesis, antioxidant, and preliminary antitumor activities of new curcumin analogues | |
| Tamura et al. | New anti-malarial phenylpropanoid conjugated iridoids from Morinda morindoides | |
| PL240095B1 (en) | 4'-O-β-D-(4"-O-Methylglucopyranosyl)-2',5'-dimethoxyflavone and method of preparing 4'-O-β-D-(4"-O-methylglucopyranosyl)-2',5'-dimethoxyflavone | |
| PL240096B1 (en) | 4'-O-β-D-(4"-O-Methylglucopyranosyl)-2',5'-dimethoxyflavone and method of preparing 4'-O-β-D- (4"-O-methylglucopyranosyl) -2', 5'-dimetoksyflavone | |
| Xie et al. | Semi‐Synthesis of Flavonoid Glycosides and Their Anti‐Inflammatory and Antitumor Activities towards Triple Negative Breast Cancer | |
| Zhang et al. | Synthesis and pharmacological activity evaluation of 2-(2-Phenylethyl) chromone analogues: The principal components in agarwood | |
| PL240947B1 (en) | 5'-O-β-D- (4"-O-Methylglucopyranosyl)-2'-methoxyflavone and method of preparing 5'-O-β-D-(4"-O-methylglucopyranosyl) -2'-methoxyflavone | |
| PL239848B1 (en) | 5'-O-β-D-(4"-O-Methylglucopyranosyl)-2'-methoxyflavone and method of preparing 5'-O-β-D-(4"-O-methylglucopyranosyl)-2'-methoxyflavone | |
| PL243174B1 (en) | 2'-O-β-D-(4"-O-Methylglucopyranosyl)-5'-methoxyflavone and method of preparing 2'-O-β-D-(4"-O-methylglucopyranosyl)-5'-methoxyflavone | |
| PL241002B1 (en) | Method of preparing 2'-O-β-D-(4"-O-methylglucopyranosyl)-flavone | |
| PL241671B1 (en) | 2'-O-β-D-(4"-O-Methylglucopyranosyl)-5'-methoxyflavone and method of preparing 2'-O-β-D-(4"-O-methylglucopyranosyl)-5'-methoxyflavone | |
| PL240957B1 (en) | 2'-O-β-D-(4"-O-methylglucopyranosyl)-flavone and method of preparing 2'-O-β-D-(4"-O-methylglucopyranosyl)-flavone | |
| PL241001B1 (en) | 5'-O-β-D-(4"-O-Methylglucopyranosyl)-2'-methoxyflavone and method of preparing 5'-O-β-D-(4"-O-methylglucopyranosyl)-2'-methoxyflavon | |
| PL240097B1 (en) | 8-O-β-D-(4"-O-methylglucopyranosyl)-2'-methoxyflavone and method of preparation 8-O-β-D-(4"-O-methylglucopyranosyl)-2'-methoxyflavone | |
| PL239565B1 (en) | 3'-O-β-D-(4"-O-methylglucopyranosyl) -flavone and method of preparing 3'-O-β-D- (4"-O-methylglucopyranosyl)-flavone | |
| PL243365B1 (en) | 2'-O-β-D-(4"-O-Methylglucopyranosyl)-5'-methoxyflavone and method of preparing 2'-O-β-D-(4"-O-methylglucopyranosyl)-5'-methoxyflavone | |
| PL239566B1 (en) | 3'-O-β-D-(4"-O-methylglucopyranosyl)-flavone and method of preparing 3'-O-β-D-(4"-O-methylglucopyranosyl)-flavone | |
| PL241672B1 (en) | Method of preparing 3'-O-β-D-(4"-O-methylglucopyranosyl)-flavone | |
| PL238787B1 (en) | 5'-Hydroxy-2'-methoxyflavone and method of preparing 5'-hydroxy-2'-methoxyflavone | |
| PL240948B1 (en) | 4'-O-β-D-(4"-O-methylglucopyranosyl)-flavone and method of preparing 4'-O-β-D-(4"-O-methylglucopyranosyl)-flavone | |
| PL240929B1 (en) | 4'-O-β-D-(4"-O-methylglucopyranosyl)-flavone and method of preparing 4'-O-β-D-(4"-O-methylglucopyranosyl)-flavone | |
| PL238786B1 (en) | 4'-Hydroxy-2',5'-dimethoxyflavone and method of preparing 4'-hydroxy-2',5'-dimethoxyflavone | |
| PL240930B1 (en) | 4'-O- -D-(4"-O-methylglucopyranosyl)-flavone and method of producing 4'-O- -D-(4'-O-methylglucopyranosyl)-flavone | |
| PL248132B1 (en) | 2'-Methyl-4'-O-β-D-(4''-O-methylglucopyranosyl)-flavone and method for preparing 2'-methyl-4'-O-β-D-(4''-O-methylglucopyranosyl)-flavone | |
| PL248131B1 (en) | 2'-Methyl-3'-O-β-D-(4''-O-methylglucopyranosyl)-flavone and method for preparing 2'-methyl-3'-O-β-D-(4''-O-methylglucopyranosyl)-flavone |