TH19482Y - A process for producing aromatic polyketide compounds that inhibit malaria pathogens from the fungus Orbiocrella. - Google Patents

Abstract

------November 27, 2020------(OCR) This invention concerns a process for producing novel aromatic polyketide compounds (I) and (II) consisting of steps involving the transplantation of fungal mycelium or spores of Orbiocrella onto a culture medium, aerobic cultivation of the fungus, fermentation of the fungus, and isolation of aromatic polyketide compounds (I) and (II) from the fungus. The compounds produced exhibit inhibitory activity against the malaria parasite (Plasmodium falciparum K1) with IC50 values of 5.7 and 5.6 micromolar, respectively. Therefore, aromatic polyketide compounds (I) and (II), or their pharmaceutical components, have the potential to be developed into antimalarial drugs (chemical formulas). ------------ Page 1 of 1. Abstract of the Invention. This invention concerns a process for producing novel aromatic polyketide compounds (I) and (II) consisting of steps involving the transplantation of fungal hyphae or spores of the fungus Orbiocrella onto a culture medium, aerobic cultivation of the fungus, fermentation of the fungus, and isolation of aromatic polyketide compounds (I) and (II) from the fungus. The compounds produced exhibit inhibitory activity against the malaria parasite (Plasmodium falciparum K1) with IC50 values of 5.7 and 5.6 micromolar, respectively. Therefore, aromatic polyketide compounds (I) and (II), or their pharmaceutical components, have the potential to be developed into antimalarial drugs (chemical formulas).

Claims (10)

------27/11/2020------(OCR) 1. The process for producing aromatic polyketide compounds (I) and (II) that exhibit inhibitory activity against the malaria pathogen (Plasmodium falciparum K1) from the fungus Orbiocrella, which consists of the following steps: a. Cultivation of Orbiocrella fungus: a.1. Transplanting of fungal hyphae or spores of Orbiocrella fungus onto the culture medium; a.2. Incubating the fungus under aerobic conditions; a.3. Fermentation of the fungus; and b. Separation of aromatic polyketide compounds (I) and (II) from the fungus, which consists of the following steps: b.1. Extraction of aromatic polyketide compounds (I) and (II) by filtering the culture medium, then extracting the culture medium (liquid) with an organic solvent. The crude extracts were selected from a group consisting of ethyl acetate, methanol, ethanol, dichloromethane, acetone, or a mixture of these substances. The appropriate organic solvent was ethyl acetate. The crude extracts contained aromatic polyketide compounds (I) and (II). B.2. Separation of aromatic polyketide compounds (I) and (II) from the crude extracts by purification using standard chromatography with an adsorbent; if normal phase silica gel was used as the adsorbent, the appropriate wash was a mixture of dichloromethane and methanol; and if reverse phase silica gel (C18) was used as the adsorbent, the appropriate wash was a mixture of acetonitrile and water; 1. The process for producing aromatic polyketide compounds (I) and (II) (chemical formulas) 2. The process for producing aromatic polyketide compounds (I) and (II) that exhibit inhibitory activity against malaria pathogens from the fungus Orbiocrella according to claim 1, in which the appropriate Orbiocrella fungus is Fungal strain Orbiocrella petchii BCC 513773. Method for the production of aromatic polyketide compounds (I) and (II) that exhibit antimalarial activity from the fungus Orbiocreiia according to claim 1, where the culture medium is potato dextrose broth.4. Method for the production of aromatic polyketide compounds (I) and (II) that exhibit antimalarial activity from the fungus Orbiocreiia according to claim 1, where the incubation steps of the fungus are in the aforementioned atmospheric conditions. This is done by shaking the inoculated fungus at a speed of 200 to 220 revolutions per minute (rpm) at a temperature of 20 to 30 degrees Celsius for 2 to 15 days.5. The process for producing aromatic polyketide compounds (I) and (II) that exhibit antimalarial activity from the fungus Orbiocreiia according to claim 1, where the fermentation of the fungus is carried out by transferring the inoculum to a suitable culture medium for secondary metabolite production. Fermentation can be carried out either in a static or shaking condition for 10 to 80 days at a temperature of 20 to 30 degrees Celsius.6. The process for producing aromatic polyketide compounds (I) and (II) that exhibit antimalarial activity from the fungus Orbiocreiia according to claim 5, where the fermentation of the fungus in the shaking condition has an optimal temperature of 25 to 28 degrees Celsius.7. The process for the production of aromatic polyketide compounds (I) and (II) with antimicrobial activity against the bacterium and fungus Orbiocreiia according to claim 1, where the purification steps by the said chromatographic method are further comprised of: — a preparative HPLC separation step using a suitable column, SunFire Prep C18 OBD, size 19 x 250 mm, silica gel size 10 µm, with a wash medium of choice from the group consisting of acetonitrile, methanol, water, or a mixture of such substances, with the appropriate wash medium being a mixture of acetonitrile and water; — a further purification step by gel filtration chromatography using Cephadex adsorbent 8. The process for the production of aromatic polyketide compounds (I) and (II) with antimalarial activity from the fungus Orbiocreiia according to claim 7, where the separation step is further purified by gel filtration chromatography using the said Sephadex adsorbent, Sephadex Trade mark LH-20, and using a wash of a choice from the group containing methanol, dichloromethane, or a mixture of such substances.9. Pharmaceutical compositions containing aromatic polyketide compounds (I), aromatic polyketide compounds (II), salts of aromatic polyketide compounds (I), salts of aromatic polyketide compounds (II), or a mixture of such substances together with pharmaceutically acceptable compounds. At least one type ------------ Page 1 of 2 Claims 1. The process for producing aromatic polyketide compounds (I) and (II) that exhibit inhibitory activity against the malaria pathogen (Plasmodium falciparum K1) from the fungus Orbiocrella consists of the following steps: a. Cultivation of Orbiocrella fungus: a.1. Transplanting of fungal hyphae or spores of Orbiocrella fungus onto the culture medium; a.2. Incubating the fungus under aerobic conditions; a.3. Fermentation of the fungus; and b. Separation of aromatic polyketide compounds (I) and (II) from the fungus, which consists of the following steps: b.1. Extraction of aromatic polyketide compounds (I) and (II) by filtering the culture medium, and then extracting the culture medium (liquid) with an organic solvent. Selected from the group consisting of ethyl acetate, methanol, ethanol, dichloromethane, acetone, or a mixture of the above substances, with the appropriate organic solvent being ethyl acetate, will obtain a crude extract containing aromatic polyketide compounds (I) and (II). b.2. Separation of aromatic polyketide compounds (I) and (II) from the above crude extract by purification by standard chromatography using an adsorbent; if normal phase silica gel is used as an adsorbent, the appropriate wash is a mixture of dichloromethane and methanol; and if reverse phase silica gel (C18) is used as an adsorbent. The appropriate detergent is a mixture of acetonitrile and water; this yields aromatic polyketide compounds (I) and (II) (chemical formulas). 2. The process for producing aromatic polyketide compounds (I) and (II) that exhibit antimalarial activity from the fungus Orbiocrella according to claim 1, in which the appropriate Orbiocrella strain is Orbiocrella petchii BCC 51377, page 2 of 2. 3. The process for producing aromatic polyketide compounds (I) and (II) that exhibit inhibitory activity against malaria-causing agents from the fungus Orbiocrella according to claim 1, where the culture medium is potato dextrose broth. 4. A method for producing aromatic polyketide compounds (I) and (II) with antimalarial activity from the fungus Orbiocrella according to claim 1, in which the incubation of the fungus under the aforementioned atmospheric conditions is carried out by shaking the inoculated fungus at a speed of 200 to 220 revolutions per minute (rpm) at a temperature of 20 to 30 degrees Celsius for 2 to 15 days. 5. A method for producing aromatic polyketide compounds (I) and (II) with antimalarial activity from the fungus Orbiocrella according to claim 1, in which the fermentation process of the fungus is carried out by transferring the inoculum to a suitable culture medium for secondary metabolite production. Fermentation can be carried out either in a static or shaking state for 10 to 80 days at a temperature of 20 to 30 degrees Celsius. 6. The process for producing aromatic polyketide compounds (I) and (II) that exhibit antimalarial activity from the fungus OrbiocreUo according to claim 5, in which the fungal fermentation under shaking conditions is carried out at an optimal temperature of 25 to 28 degrees Celsius. 7. A procedure for the production of aromatic polyketide compounds (I) and (II) with antimicrobial activity against the fungus OrbiocreUo according to claim 1, in which the purification procedure by chromatography is further comprised of: — a preparative HPLC separation step using a suitable column, namely SunFire Prep C18 OBD, size 19 x 250 mm, silica gel size 10 µm, with a wash medium of choice from the group consisting of acetonitrile, methanol, water, or a mixture of such substances, with the appropriate wash medium being a mixture of acetonitrile and water; — a further purification step by gel filtration chromatography using Cephadex adsorbent. 8. A method for the production of aromatic polyketide compounds (I) and (II) with antimalarial activity from the fungus OrbiocreUo according to claim 7, where the separation step is further purified by gel filtration chromatography using the said Sephadex adsorbent, Sephadex®LH-20, and a wash of a choice of a group containing methanol, dichloromethane, or a mixture of the above. 9. Pharmaceutical compositions containing aromatic polyketide compounds (I), aromatic polyketide compounds (II), salts of aromatic polyketide compounds (I), salts of aromatic polyketide compounds (II), or a mixture of such compounds, together with at least one pharmaceutically acceptable compound. 10. A method for inhibiting the malaria parasite (Plasmodium falciparum K1) in living organisms, in which the method comprises steps for delivering the pharmaceutical component as described in claim 9 into the body of the organism.